CN105310923B - The manufacturing method of Australia La Woer chrysanthemum young shoot extract of cosmetic composition and tissue cultures - Google Patents
The manufacturing method of Australia La Woer chrysanthemum young shoot extract of cosmetic composition and tissue cultures Download PDFInfo
- Publication number
- CN105310923B CN105310923B CN201510232464.XA CN201510232464A CN105310923B CN 105310923 B CN105310923 B CN 105310923B CN 201510232464 A CN201510232464 A CN 201510232464A CN 105310923 B CN105310923 B CN 105310923B
- Authority
- CN
- China
- Prior art keywords
- australia
- woer
- tissue cultures
- young shoot
- chrysanthemum
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q1/00—Make-up preparations; Body powders; Preparations for removing make-up
-
- 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
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—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
Abstract
The present invention relates to the cosmetic compositions and its manufacturing method of Australia La Woer chrysanthemum young shoot (tissue cultured Raoulia australis shoot) extract comprising tissue cultures; Australia La Woer chrysanthemum extract of tissue cultures of the invention is due to excellent anti-oxidant, anti-inflammatory and whitening effect, can usefully be used in the cosmetic composition for the purpose of skin sparing and skin improvement.
Description
Technical field
The present invention relates to a kind of cosmetic composition and its manufacturing methods, and in particular to anti-oxidant, anti-inflammatory and whitening effect
The cosmetic composition and its manufacturing method of excellent Australia La Woer chrysanthemum young shoot extract comprising tissue cultures.
Background technique
Skin has the function of protecting the body from outside stimulus.This outside stimulus, that is, environment, physics, chemical factor with
Internal factor physiologically not only reduces skin function, compared with normal skin, causes scytitis to outside stimulus and such as mole
The reason of with the pigmentation disease of spot etc, also reduction skin function as skin aging.
Therefore present cosmetics exploitation in constantly look for cutaneous safety is excellent, can protect the skin from it is various because
Element influences, anti-inflammatory, anti-oxidant and whitening effect etc. can improve and protect the stage of the cosmetics new material of skin.
It is representative to have solubilizer, cream in constituting the ingredient in order to protect the cosmetics of skin, beautification cleaning and exploitation
Agent, preservative, fragrance, UV blockers, synthesis tackifier etc., but these ingredients be generally believed that be scytitis or
The induced factor of acne etc..In addition, the fatty acid, protein etc. in the sebum generated in vivo, sweat and cosmetic composition can be with skins
Various bacterium present in skin react and induce inflammation.
The intracorporal active oxygen of people is generated by the reaction of various enzymes, to the biosynthesis of physiological activator, immune function,
Drug metabolism etc. plays a very important role.But if not maintaining internal shape constancy, when generating excessive, human body is damaged instead
Wound.For example, free radical and active oxygen make the body materials such as Cell membrane lipids, protein, nucleic acid aoxidize or go bad, skin is reduced
The function of cell, thus the main reason for becoming skin problem, inflammation, skin aging.
There are many kinds of the factors for determining human body skin color, wherein important has melanocyte (melanocyte), skin
Skin thickness, the distribution of blood vessel and whether contain the factors such as pigment inside and outside human body.Especially most important factor is intracorporal in people
The black pigment for being referred to as melanin generated in melanocyte by the various enzyme effects such as tyrosinase.Melanin pigment is easily logical
Cross the skin pigmentation disorders calmness symptoms such as mole, freckle, the pigmentation generated by the external environmental factor of ultraviolet light irradiation etc
Deng and formed.Calm in order to treat or mitigate excessive melanin pigment, various substances are used cooperatively with cosmetics or drug.But
Be, due to whitening effect it is insufficient, to safety issue, the cosmetic ingredients of skin when dosage form and stability problem etc.,
Its use is restricted.
The effort of beautiful skin of preventing from keeping fit while these skin phenomenas is also continuing.Skin is improved recently
Research direction be exploitation it is without side-effects, with human body is affine, can independently feel at ease with constitution using utilize plant extracts
Material.
In general, the tropical rain forest gas that can be grown in plant can be used in the function cosmetics comprising natural plant extracts
Time, subfrigid zone, all plants grown in climate of frigid zone.Plant adapts to respective weather, with best biological machine under this condition
System growth.Therefore, even kindred plant, according to according to its sunshine amount, temperature, precipitation of latitude for growing etc. and soil acid
Degree, it is fertile degree etc. weather conditions and Various Seasonal, plant different parts, 2 metabolites of inside plants etc it is effective
The type of ingredient is different from content.Moreover, it is difficult that supply is overcome to be difficult to meet manufacture, the disadvantages of processing charges is more.In order to
Improve these all difficulties, it is significant using the cosmetic material development effectiveness of vegetable culture technique recently.
On the other hand, wild herb Australia La Woer chrysanthemum (Raoulia australis) of composite family is wild is grown in
New Zealand and Australia.La Woer chrysanthemum is grown on the humus soil easily drained with mat form in Australia, due to cold resistance
By force, main wild is grown in the cobble of alpine belt, rock surface, always using ground cover plant when being Landscape.Australia
The substance for the terpene series that the La Woer sour (Raoulic acid) for drawing Wall chrysanthemum to contain is made of 25 carbon, research shows that it has
Have to the virus for causing paranasal sinusitis, tympanitis, chronic bronchitis, aseptic meningitis, acute pancreatitis, myocarditis etc.
Proliferation inhibiting effect.
Have effects that these Australia La Woer chrysanthemum should not be cultivated in the country of South Korea due to geographical location and climate change.
It is known to tissue cultures although increase in demand finds various effects by research as effective component is realized
The scientific experiment result of the effect of Australia La Woer chrysanthemum young shoot is seldom.In addition, being also grinding for cosmetic composition without any application
Study carefully the report of result.
Summary of the invention
The object of the present invention is to provide a kind of cosmetic composition and its manufacturing method, in the cosmetic composition
It draws in Australia of tissue cultures excellent comprising anti-oxidant, anti-inflammatory and whitening effect and with skin improvement and skin sparing effect
The extract of Wall chrysanthemum young shoot (tissue cultured Raoulia australis shoots).
The purpose of the present invention is not limited to the above-mentioned purpose referred to, and unmentioned other purposes can be defined from following records
Ground understands.
Cosmetic composition according to the present invention, which is characterized in that Australia La Woer chrysanthemum young shoot containing tissue cultures
(tissue-cultured Raoulia australis shoots) extract is effective component.
It herein, preferably include described with the range of 0.1~10 weight % relative to the cosmetic composition total weight
Australia La Woer chrysanthemum young shoot extract of tissue cultures.
In particular, Australia La Woer chrysanthemum young shoot extract of the tissue cultures, which can be used, is selected from pure water, the alcohol of C1~C4,
And the solvent extraction tissue cultures of one or more of polyalcohol of C1~C5 Australia La Woer chrysanthemum young shoot and obtain.
Also, cosmetic composition according to the present invention can have selected from skin cream, smoothing toner, toner, astringent lotion,
Lotion, milky lotion, nutritional emulsions, massage cream, nourishing cream, hand lotion, vanishing cream, sun screen, dual-purpose muffin, mascara, essence
Dosage form in China, nutrition essence, facial mask, soap, skin cleaning foam, clean skin dew, cleansing cream, body lotion and shower cream.
In addition, the manufacturing method of Australia La Woer chrysanthemum young shoot extract of tissue cultures according to the present invention, may include:
(a) by Australia La Woer chrysanthemum young shoot of tissue cultures in the polyalcohol of alcohol and C1~C5 selected from pure water, C1~C4
The step of being impregnated 3~30 days in more than one Extraction solvent and forming dipping solution；And (b) filter the dipping solution and
The step of obtaining Australia La Woer chrysanthemum young shoot extract of tissue cultures.
Herein, the Extraction solvent can be used mixed solvent selected from ethyl alcohol, ethyl alcohol and 1,3-BDO, ethyl alcohol with
Any one of mixed solvent and ethyl alcohol and the mixed solvent of pentanediol of propylene glycol.
In addition, Australia La Woer chrysanthemum young shoot weight relative to the tissue cultures, the Extraction solvent preferably have 10
~20 times of weight ratio.
Cosmetic composition according to the present invention includes Australia La Woer chrysanthemum young shoot extract of tissue cultures, will not be stimulated
Skin has excellent anti-oxidant, anti-inflammatory and whitening effect, and highly effective to skin improvement and skin sparing.Tissue training
Feeding Australia La Woer chrysanthemum young shoot (tissue cultured Raoulia australis shoots) extract protects skin
Not by various outside stimulus, it is moist to make skin slip, and shows the effect of the highly significant improved to skin function.
Specific embodiment
The present invention can carry out numerous variations, and can have various embodiments, exemplify specific embodiment, and by detailed
Illustrate that this is described in detail.But this is not to limit the invention to specific embodiment, it is thus understood that is included in
Include in thought and technical scope of the invention have altered, equipollent to substitute.
In the following, to Australia La Woer chrysanthemum young shoot (tissue cultured according to the present invention comprising tissue cultures
Raoulia australis shoots) cosmetic composition of extract is illustrated.
The present inventor is with Australia La Woer chrysanthemum young shoot (tissue cultured Raoulia to tissue cultures
Australis shoots) extract the effect of effect evaluation centered on, continue to using vegetable culture technique as change
The application possibility of cosmetic is studied, and then completes the present invention.As a result, the present inventor specifies that tissue is trained
Feeding Australia La Woer chrysanthemum young shoot (tissue cultured Raoulia australis shoots) extract protects skin
Not by various outside stimulus, keep skin slip moist, and has the effect of highly significant to skin function improvement.
Relative to the total weight of composition, cosmetic composition according to the present invention is preferably with the model of 0.1~10 weight %
Enclose the extract of Australia La Woer chrysanthemum young shoot comprising tissue cultures.Helpfulness effect can be showed by meeting the range, that is, be protected
Sheath skin by various outside stimulus, it is moist not make skin slip, and shows the effect for facilitating skin function improvement.Separately
Outside, it is contemplated that intrinsic smell, color of the extract of final products etc. are preferably included as the range.
The extract of Australia La Woer chrysanthemum young shoot of the tissue cultures it is preferable to use selected from pure water, C1~C4 alcohol,
And the solvent of one or more of polyalcohol of C1~C5 extracts Australia La Woer chrysanthemum young shoot of tissue cultures.
The product of cosmetic composition of the invention can be added, such as has various skin creams, smoothing toner, toner, receipts
Hold back water, lotion, milky lotion, nutritional emulsions, massage cream, nourishing cream, hand lotion, vanishing cream, sun screen, dual-purpose muffin, eyelashes
Cream, essence, nutrition essence, facial mask, soap, skin cleaning foam, clean skin dew, cleansing cream, body lotion and shower cream etc., but and it is unlimited
Due to this.It, can be containing to the formulation of the dosage form when cosmetic composition of the invention is manufactured into the dosage form of the form
Various matrix required and appropriate and additive.In addition, the type of these ingredients and amount can be easy the technology belonging to the present invention
The technical staff in field is selected.
In the following, illustrating the manufacturing method of Australia La Woer chrysanthemum young shoot extract of tissue cultures of the invention.
The present invention include: (a) by Australia La Woer chrysanthemum young shoot of tissue cultures selected from pure water, C1~C4 alcohol, with
And the step of being impregnated 3~30 days in the Extraction solvent of one or more of polyalcohol of C1~C5 and forming dipping solution；And
(b) the step of filtering the dipping solution and obtaining Australia La Woer chrysanthemum young shoot extract of tissue cultures.
Australia La Woer chrysanthemum young shoot of above-mentioned tissue cultures used in (a) step is preferably drying regime.And
The dip time of (a) step is preferably 10~20 days.
The weight of Australia La Woer chrysanthemum young shoot of tissue cultures relative to the drying, the Extraction solvent is preferably with 10
~20 times of weight ratio (w/w) uses.
The alcohol of the C1~C4 can be methanol, ethyl alcohol, propyl alcohol, butanol etc., particularly preferably ethyl alcohol (Ethyl
alcohol).In addition, the polyalcohol of the C1~C5 is preferably selected from alkylene glycol, 1,3-BDO, propylene glycol and pentanediol
One or more of.
In addition, mixing ratio (w/w) is preferably 6 when the Extraction solvent is the mixed solvent of the alcohol and alkylene glycol:
(that is, the total weight of whole mixed solvents relative to pure and mild alkylene glycol, the content of alcohol is preferably 60~90 weights to 4~9:1
Measure %).The content of the mixed solvent of the alcohol and alkylene glycol has to effective component needed for only extracting more in the range
Effective polarity.Herein, the concentration of alcohol used in the mixed solvent of the alcohol and alkylene glycol be preferably 60%~
100%.
In addition, the Extraction solvent may be the mixed solvent of the pure water Yu the alkylene glycol, mixing ratio
Preferably 3:7~1:9 is (that is, the total weight relative to pure water and whole mixed solvents of alkylene glycol, the content of alkylene glycol
Preferably 70~90 weight %).
Preferably ethyl alcohol when the Extraction solvent is single solvent.Also, Extraction solvent can be used when being mixed solvent
The mixed solvent of ethyl alcohol and alkylene glycol is, it is preferable to use ethyl alcohol and 1,3-BDO, ethyl alcohol and propylene glycol or ethyl alcohol and penta 2
The mixing of the mixed solvent or ethyl alcohol and pentanediol of ethyl alcohol and 1,3-BDO more preferably can be used in the mixed solvent of alcohol
Solvent.
In addition, (b) step preferably further includes the steps that removing Extraction solvent.
Present invention offer includes that Australia La Woer chrysanthemum young shoot extract of the tissue cultures obtained by the manufacturing process is
The cosmetic composition of effective component.Herein, the Australia of tissue cultures according to the present invention is confirmed by following steps
The effect of continent La Woer chrysanthemum young shoot extract, wherein include: Australia La Woer chrysanthemum young shoot extract pair with the tissue cultures
The step of human fibroblasts are handled, and the cytotoxic effect to skin irritatin is investigated；With Australia of the tissue cultures
The step of drawing Wall chrysanthemum young shoot extract to handle human fibroblasts, investigating the cytoprotective effect to outside stimulus；
By Australia La Woer chrysanthemum young shoot extract to the tissue cultures carry out lipoxygenase (Lpoxgenase) activity suppression and
The step of generation rejection ability of nitrogen oxide is tested, and antiphlogistic effects are investigated；Pass through (the SUPEROXIDE of superoxide anion
ANION the step of) elimination activity and free radical (HYDROXY RADICAL) elimination activity ability, investigation antioxidant effect；Investigation
Tyrosinase inhibits the step of bring skin-whitening improvement.
Moreover, cosmetic composition according to the present invention is not limited to dosage form exemplified below, can be led using this technology
Dosage form known to domain.Cosmetic composition of the invention can be in the range of not damaging the purpose of the present invention and effect in institute
The other compositions usually cooperated in cosmetics can be cooperated by stating in extract.
Production Example, test example and dosage form example according to the present invention are enumerated below and carries out more specific description, but these are
In order to illustrate the present invention, it's not limited to that for the scope of the present invention.
[1~Production Example of Production Example 4 and 1~comparison manufacturing example of comparison manufacturing example 4]: Australia La Woer of tissue cultures
The manufacture of chrysanthemum young shoot extract
Australia La Woer chrysanthemum young shoot of tissue cultures and natural Australia La Woer chrysanthemum are washed respectively, remove water with gauze
Afterwards, dry and chopping.Herein, " Australia La Woer chrysanthemum young shoots of tissue cultures " used in this Production Example are learned by loyal Beijing University
Tip horticulture technique developmental research center provide, it be by from Australia La Woer chrysanthemum seed of New Zealand's import in sterile item
Young shoot is induced using plant tissue culture technique under part, made of Solution culture method.
By Australia La Woer chrysanthemum young shoot of ready tissue cultures and Australia La Woer chrysanthemum in listed in Table 1 content
Existing in the mixed solvent impregnates 2 weeks, is cured.Thereafter with filter paper (Japanese ToyoroshiKaisha, Ltd. sale 5C,
It 185mm) filters, ethanol evaporation, obtains Australia La Woer chrysanthemum young shoot extract and Australia La Woer of the tissue cultures of filtrate shape
Chrysanthemum extract.
[table 1]
(b1:70% ethyl alcohol/b2:1,3- butanediol/b3: propylene glycol)
The cytotoxicity assay of Australia La Woer chrysanthemum young shoot extract of [test example 1] tissue cultures
By the cell toxicity test to Production Example 1 and comparison manufacturing example 1, safety is had rated.
[cell culture (Cell culture)]
By from the CCD-1064sk human fibroblasts that ATCC company buys in 10cm Tissue Culture Dish, to be added with
The Yi Sikefu improvement culture of 10% fetal calf serum (FBS), 1% Pen .- Strep (penicillin-streptomycin)
Liquid (Iscove's modified Dulbeco's medium:IMDM) 10ml be culture medium cultivated so that 37 DEG C,
5%CO2Cell fusion (confluency) 70%~80% in incubator.Culture medium is replaced weekly 2 times, and the cell of fusion is reached
After carrying out pancreatin processing (trypsinization) using pancreas enzyme -EDTA (trypsin-EDTA), maintained with secondary culture.By institute
The human fibroblasts of culture are stated respectively with 1 × 104Cells/well point cultivates 24 on 96 orifice plates (96-multiwell plate)
Hour.With the culture medium of the cell of 2%FBS culture medium replacement culture, with concentration for the treatment of respectively with culture medium dilution Production Example 1
After Australia La Woer chrysanthemum young shoot extract of tissue cultures and Australia La Woer chrysanthemum extract of comparison manufacturing example 1, in CO2Culture
It is cultivated 48 hours in case.
[test example 1-1]: the measurement of cytotoxic effect (MTT detects (assay))
The MTT that PBS (phosphate buffered saline (PBS) (phosphate buffered saline)) will be dissolved in is saved
(stock) solution (5mg/ml), which is added in new culture medium, dilutes 10 times, its solution is added 100 μ l respectively and is trained to end is placed with
In the hole of feeding cell, reacted 4 hours at 37 DEG C.Because of dehydrogenase and yellow water in the mitochondria of the cell to live after reaction
Soluble base MTT [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium-bromide (3-
(4,5- dimethyl -2- thiazolyl) -2,5- diphenyl -2H- tetrazoleBromide)] it is reduced to water-insoluble MTT- first(MTT-formazan) it crystallizes, the survival rate of cell can be measured by detecting its amount.Culture supernatant is removed, in each Kong Zhongtian
200 μ l of DMSO is added to dissolve the MTT- first generated in cellAfter crystal, with absorb spectrophotometer (UVmax,
Molecular Device, USA) measurement can make firstThe maximum test optical filter of absorbance (Test filter)
The O.D value of 540nm wavelength and the O.D value of basic filter (Reference filter) 630nm wavelength, find out suction with its difference
Luminosity (bibliography The EMBO Journal (2002) 21,2407-2417 is outer most).Cytotoxicity is with the percentage of absorbance
Than showing.
※ cells survival rate (%)=(sample pretreating group absorbance/non-treated group of absorbance) × 100
Being averaged for non-treated group of absorbance and the absorbance of sample pretreating group is calculated based on original (Raw) data
Value calculates standard deviation.
[table 2]
[test example 1-2]: the measurement of cytotoxic effect (dimethyl diaminophenazine chloride detects (Neutral Red assay))
Dimethyl diaminophenazine chloride (Neutral Red, Sigma-Aldrich, USA) is separately added into each hole for the cell that culture terminates
50 μ g/ml react 3 hours.Dimethyl diaminophenazine chloride be concentrated in the cell survived completely or the cytase body that does not damage in.After reaction,
With 1.0% formalin/1.0% chlorination Potassium fixating cell, 1.0% acetic acid/50% ethanol solution is then added, is extracted neutral
It is red.By the dimethyl diaminophenazine chloride of extraction using absorb spectrophotometer (UVmax, Molecular Device, USA) measurement 540~
Each levels of cytotoxic value of 630nm, and it is shown in following Table 3.Cytotoxicity is shown with the percentage of absorbance, with 3 batches (lot)
Implement 3 times.
※ cells survival rate (%)=(sample pretreating group absorbance/non-treated group of absorbance) × 100
[table 3]
Being averaged for non-treated group of absorbance and the absorbance of sample pretreating group is calculated based on original (Raw) data
Value calculates standard deviation.
As shown in table 2 and table 3, the MTT of manufacture grain 1 of the invention50、NR50Value is 1% or more, it is known that with comparison manufacturing example
1 compared to being safe raw material.
[test example 2]: the cytoprotective effect measurement of Australia La Woer chrysanthemum extract of tissue cultures
Carry out SDS (lauryl sodium sulfate (the Sodium dodecyl to Production Example 1 and comparison manufacturing example 1
Sulfate)) the cytoprotective effect experiment of cytotoxicity.
With the IMDM added with 10% fetal calf serum (FBS), 1% Pen .- Strep in 37 DEG C, 5%CO2In incubator
Cultivate human fibroblasts.By the human fibroblasts of culture with 1 × 104Cells/well is divided respectively in 96 orifice plates, and culture 24 is small
When.With the culture medium of the cell of 2%FBS culture medium replacement culture, by SDS (Sodium dodecyl sulfate) 0.002%
After pre-processing 1 hour, Production Example 1 and comparison manufacturing example 1, parallel processing are diluted respectively with culture medium with respective concentration for the treatment of
Afterwards, in CO2It is cultivated 24 hours in incubator.10 times of dilutions are dissolved in PBS (phosphate buffered in new culture medium
Saline the MTT in) saves solution (5mg/ml), its solution is added each 100 μ l to each of the cell for having terminated culture respectively
From in hole, reacted 4 hours at 37 DEG C.Because of dehydrogenase and yellow water-soluble matrix in the mitochondria of the cell to live after reaction
MTT [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium-bromide] is reduced to non-aqueous
The MTT- first of property(MTT-formazan) it crystallizes, the survival rate of cell can be measured by detecting its amount.Culture supernatant is removed,
200 μ l of DMSO is added in each hole to dissolve the MTT- first generated in cellAfter crystal, to absorb spectrophotometer
(UVmax, Molecular Device, USA) measurement can make firstThe maximum test optical filter 540nm wavelength of absorbance
O.D value and basic filter 630nm wavelength O.D value, absorbance is found out with its difference.(bibliography The EMBO
Journal (2002) 21,2407-2417 is outer most).Cytoprotective effect is shown with the percentage of absorbance.
Cells survival rate (%)=(sample pretreating group absorbance/non-treated group of absorbance) × 100
Being averaged for non-treated group of absorbance and the absorbance of sample pretreating group is calculated based on original (Raw) data
Value calculates standard deviation.
[table 4]
As shown in table 4, it is known that, for outside stimulus, the cytoprotective effect and comparison manufacturing example of Production Example 1 of the invention
1 compares, and can more promote cytoprotective effect.
[test example 3]: the measurement of the antiphlogistic effects of Australia La Woer chrysanthemum young shoot extract of tissue cultures
Measurement Production Example 1 and comparison manufacturing example 1 carry out antiphlogistic effects to lipoxygenase activity and nitric oxide production quantity
Experiment.
Inflammatory reaction is human body to maintain shape constancy, is reacted the biophylaxis of outside stimulus.Caused by generating stimulation
The approach of the enzyme of inflammation is using lipoxygenase activity approach and the active approach of cyclooxygenase (cycloxygenase) as representative.Rouge oxygen
Synthase is catalyzed the hydroperoxidation (hydroperoxidation) of unsaturated fatty acid, generates lipid peroxide hydrogen (fatty
Hydroperoxides), the decomposition products such as these substances regeneration epoxides (epoxide), hydroxyl, ketone, aldehyde.Lipoxygenase
From by linolenic acid (linolenic acid) biosynthesis arachidonic acid (arachidonic acid) generate and organism
Relevant a variety of chemical mediators (chemical mediator) such as interior inflammatory reaction or allergy.Therefore, the object of lipoxygenase is hindered
Matter shows the knot for inhibiting the generation of chemical mediator (chemical mediators) relevant to various inflammatory reactions or allergy etc.
Fruit.Although by the nitric oxide (NO) generated using inflammatory reaction as the activation of the enzyme of medium etc. in immune response it is important
Effect, but excessive nitric oxide can cause inflammatory reaction, bring exception to immune system.
[test example 3-1]: the measurement of the generation rejection ability of nitrogen oxide
The generation degree of nitrogen oxide (NO) is with method (Green LC, Wagner DA, the Glogowski J, et of Green etc.
al.1982.Analysis of nitrate,nitrite,and[15N]nitrate in biological fluids.Anal
Biochem 126:131-138.) amount of nitrate that generates in culture medium of the measurement as nitrogen oxide (NO) generation index, into
Row analysis.That is, processing sample adds LPS (lipopolysaccharide, 1 μ g/mL) after culture 30 minutes, cultivate 24 hours
And Raw264.7 macrophage-conditioned media is obtained, in 100 μ L medium supernatants of the Raw264.7 macrophage-conditioned media
Add 1% sulfanilamide (SN) (being dissolved in the sulfanilamide (SN) (sulphanilamide) in 5% phosphoric acid (phosphoric acid)) and 50 of 50 μ L
The 0.1% naphthylenediamine dihydrochloride (naphthylenediamine dihydrochloride) of μ L, in room temperature reaction 10 minutes
Afterwards, absorbance is measured in 540nm.Control group for the anti-inflammatory activity effect test uses the experiment of individually processing LPS
Group.
※ NO inhibiting rate (%)=(the NO amount of control group-each extract NO amount/control group NO amount) × 100
Average value is calculated based on original (Raw) data, finds out standard deviation.
[table 5]
The generation rejection ability of the NO occurred in oxidative stress caused by LPS result as shown in the table 5, manufacture
Example 1 shows that about 1.5~2 times of NO generates inhibitory effect compared with comparison manufacturing example 1.
[test example 3-2]: the activity suppression of measurement lipoxygenase (Lipoxygenase)
Implement the active suppression test of the lipoxygenase of the representative enzyme as the inflammation forming factors to outside stimulus.
Mixed into 1mM linolenic acid (Sigma-aldrich, USA) 1ml sample to be tested 0.1ml and lipoxygenase (Sigma-aldrich,
USA) 0.9ml after carrying out whirlpool concussion (voltexing), reacts 10 minutes at 25 DEG C.20% trichloroacetic acid is then added
(trichloroacetic acid) 0.5ml and 0.6% thiobarbituricacidα- (thiobarbituric acid) 1ml, in hot water
After ten minutes, cold soaking 2-3 minutes in ice water, butanol (butanol) 2ml is added in middle reaction, after whirlpool concussion 20 seconds or more, with
3500rpm is centrifugated 5 minutes, O.D value is measured in 513nm with UV spectrophotometer (spectrophotometer), under
The maximum inhibition that mathematical expression 2 calculates lipoxygenase is stated, is shown in table 6.Measurement test point 3 batches (batch) implements 3 times,
The average value that absorbance is calculated based on original (Raw) data, finds out standard deviation.
[table 6]
As shown in table 6, Australia La Woer chrysanthemum young shoot extract of the tissue cultures of Production Example 1 and non-treated control group ratio
Compared with when, rely on concentration inhibit lipoxygenase, be able to confirm that compared with comparison manufacturing example 1, the activity suppression to the enzyme for causing inflammation
Effect is outstanding.
The antioxidant effect of Australia La Woer chrysanthemum young shoot extract of [test example 4] tissue cultures measures
The antioxidant effect for implementing Production Example 1 and comparison manufacturing example 1 is tested.
[test example 4-1] DPPH (1,1- diphenyl -2- picryl hydrazine (1,1-diphenyl-2picrylhydrazyl)) is certainly
Anti- hydroxyl radical free radical active (%) measurement as caused by base
It is radical reaction that reaction caused by active oxygen is almost most of, includes Auto-oxidation reaction process.In organism
Interior antioxidant is responsible for blocking the effect of Auto-oxidation reaction.Reducing power as electron donation is stronger, can more become strong
Antioxidant.The reagent for measuring the reducing power of antioxidant has 1,1- hexichol -2- picryl hydrazine (1,1-diphenyl-2-
Picrylhydrazyl:DPPH) free radical.The free radical form of rock-steady structure of the DPPH in compound centered on nitrogen is deposited
?.Absorption maximum is shown in 517nm, and the absorption for being reduced rear 517nm disappears.DPPH (D9132, SIGMA) is dissolved in ethyl alcohol
With the in the mixed solvent of the 6:4 (v/v) of distilled water, the absorbance 0.6 or so that the DPPH solution makes it in 513nm is adjusted.
According to DPPH solution 2ml is added in the sample 1ml of concentration dilution respectively, mixed.After placing about 1 hour under room temperature, make
With the absorbance for absorbing spectrophotometer (UVmax, Molecular Device, USA) measurement 513nm, table the results are shown in
7。
[table 7]
As shown in table 7, Production Example 1 shows excellent antioxidant effect.
[test example 4-2]: anti-hydroxyl radical free radical active (%) measurement caused by ABTS free radical
Utilize the total antioxidant capacity of the method measurement extract of Miller [Miller et al. (1993)].With the dense of 7mM
It spends bis- (3- ethyl benzo thiazole phenanthroline -6- sulphonic acid ester) (2,2'-Azino-bis (3-ethyl of 2,2'- azine group -
Benzothiazoline-6-sulfonate it)) is dissolved in distilled water, was added in such a way that ultimate density is 2.45mM thereto
Cationic (cation) free radical of ABTS is made in potassium sulfate (Potassium persulfate).By ABTS and potassium peroxydisulfate equivalent
Mixing, is placed at room temperature in the dark after 12 hours, this solution is diluted in the ethyl alcohol (ethanol), adjust to its 734nm extinction
Degree is 0.7.10 μ l of mixed plant extract sample, 190 μ l of ABTS solution (Solution), 50 μ l of distilled water are dividing after 6 minutes
Light photometer (spectrophotometer) 734nm measures absorbance, finds out absorbance difference with percentage (%), is tied
Fruit is shown in table 8.
Elimination factor (%)=(B-A)/B × 100
(B: the absorbance of the non-addition group of extract, A: the absorbance of extract addition group)
[table 8]
As shown in table 8, Australia La Woer chrysanthemum extract (Production Example 1) of tissue cultures according to the present invention frequently relatively manufactures
Example 1 shows better antioxidant effect.
[test example 5]: the tyrosinase activity and B16 cell of Australia La Woer chrysanthemum young shoot extract of tissue cultures
Inhibitory effect measurement.
[test example 5-1] tyrosinase activity hinders test
In order to investigate the whitening active of Production Example 1 and comparison manufacturing example 1, tyrosinase activity is carried out and has hindered test.For
Test, with phosphate buffer (pH6.5) dissolution Mushroom Tyrosinase (mushroom tyrosinase, T-3824,
1530U/mg, Sigma) enzyme solution to 1000U/ml as tyrosinase, dissolves l-tyrosine with phosphate buffer (pH6.5)
(L-tyrosine, 45160-0410, Junsei chemical co.Ltd) is to 1.5mM as matrix liquid.
Production Example 1 and comparison manufacturing example 1 are diluted with various concentration with buffer, prepared detection liquid.Detection liquid 170 is added
The 10 μ l of enzyme solution of μ l, tyrosinase are placed 10 minutes at 37 DEG C.After 20 μ l of matrix liquid wherein is added, reacted 10 minutes at 37 DEG C
Afterwards, it places in ice 5 minutes, is then measured using absorption spectrophotometer (UVmax, Molecular Device, USA) immediately
The absorbance of wavelength 490nm.The absorbance of the measurement is substituted into following mathematical expressions, the activity for calculating tyrosinase hinders
Rate is shown in table 9.Substitution detection liquid is put into phosphate buffer (pH6.5) and the liquid of manufacture substitutes base as blank test liquid
Matter liquid is put into phosphate buffer (pH6.5) and the liquid of manufacture is as opaquing fluid.
<mathematical expression>
Tyrosinase activity obstruction rate (%)=100- (A-A')/(B-B') × 100
A: the absorbance after detection liquid reaction
B: the absorbance after the reaction of blank test liquid
A': the opaquing fluid of liquid is detected
B': the opaquing fluid of blank test liquid
[table 9]
[test example 4-2]: inhibited according to the B16 cell of Australia La Woer chrysanthemum young shoot extract concentrations of tissue cultures
Measurement experiment
To Production Example 1 and comparison manufacturing example 1, B-16 melanocyte (Melanoma cell: Korea Cell strain silver is utilized
Row), measure whitening effect.Measuring method be will from black mouse B-16 melanocyte with containing 10%FBS (GIBCO.,
USA DMEM) is with 2 × 104The concentration of cells/well respectively adds 2ml to 6 hole tissue culture dishes, in 5%CO2, train under the conditions of 37 DEG C
It supports 24 hours.Culture medium is removed after culture, to contain 10%FBS, 2uM a-MSH (Sigma., USA, melanophorin
(Melanocyte stimulating Hormone)) it is replaced with the DMEM of 2mM theophylline (Sigma., USA, theophylline)
Afterwards, after adding Australia La Woer chrysanthemum young shoot extract with the suitably diluted tissue cultures of same medium respectively, in 5%CO2、
Culture reaches about 80% or more in bottom hole to cell under the conditions of 37 DEG C.Culture medium is removed after culture, then with PBS (Sigma.,
USA, Phosphate buffered Saline) washing, after handling with pancreatin, recycle cell.The cell of recycling is with 5000rpm
Centrifuge separation removes supernatant after ten minutes.After drying 24 hours, 1N is added in 60 DEG C of thermostats in the cell for removing supernatant
NaOH dissolves intracellular melanin.Using absorption spectrophotometer (Uvmax, Molecular Device, USA) in wavelength
The absorbance of the melanin of 490nm measurement dissolution.
<mathematical expression>
Melanin production obstruction rate (%)=(1-A/A') × 100
(A: test group absorbance, A': control group absorbance)
[table 10]
As shown in table 9 and table 10, inhibit to 1 concentration dependent of Production Example the activity of tyrosinase, and than comparison manufacturing example 1
Inhibitory effect is generated with superior intracellular melanin.
The essence of Australia La Woer chrysanthemum young shoot extract of [1~embodiment of the embodiment 2] manufacture containing tissue cultures
The essence comprising Production Example 1 has been manufactured according to the composition of following table 11.
[table 11]
In table 11, fully dispersed, moistening ingredients A forms uniform gel (Gel) shape, and addition B is neutralized.Ingredient (A+
B ingredient C is added in), after uniform stirring dissolution, ingredient D is added at room temperature, stirring is uniformly distributed it, contains complete in container
At commercialization.
[embodiment 3~4]: the soft and moist toner (skin of Australia La Woer chrysanthemum young shoot extract of the manufacture comprising tissue cultures
Skin)
The embodiment of soft and moist toner (skin) is as shown in table 12 below in cosmetic composition comprising Production Example 1.
[table 12]
[embodiment 5~6]: the frost of Australia La Woer chrysanthemum young shoot extract of the manufacture comprising tissue cultures includes Production Example 1
Cosmetic composition in white prescription for example shown in the following table 13.
[table 13]
Although until now, preferred embodiments of the present invention have been disclosed for illustrative, the technology of the technical field of the invention
Personnel can realize the form of deformation in the range of without departing from intrinsic propesties of the invention.Therefore, the present invention described herein
The viewpoint that does not limit of embodiment, considered that the scope of the present invention is embodied in claim from the viewpoint of explanation
In range, rather than in the explanation, it should be interpreted that the present invention includes all discrepancys in its equivalents.
Claims (6)
1. a kind of cosmetic composition, which is characterized in that Australia La Woer chrysanthemum (Raoulia comprising tissue cultures
Australis Australia La Woer chrysanthemum young shoot extract of) young shoot extract, the tissue cultures is using ethyl alcohol and 1,3- fourth two
The mixed solvent or ethyl alcohol of alcohol and the mixed solvent of propylene glycol extract made of Australia La Woer chrysanthemum young shoot of tissue cultures.
2. cosmetic composition according to claim 1, which is characterized in that relative to the composition total weight, with 0.1
The range of~10 weight % includes Australia La Woer chrysanthemum young shoot extract of the tissue cultures.
3. cosmetic composition according to claim 1, which is characterized in that the cosmetic composition, which has, is selected from moisturizing
Dew, smoothing toner, toner, astringent lotion, lotion, massage cream, nourishing cream, hand lotion, vanishing cream, sun screen, dual-purpose muffin, eyelashes
Dosage form in cream, essence, facial mask, soap, skin cleaning foam, clean skin dew, cleansing cream and shower cream.
4. cosmetic composition according to claim 3, which is characterized in that the cosmetic composition, which has, is selected from milk
The dosage form of lotion, nutritional emulsions, body lotion and nutritious extract Central China.
5. a kind of manufacturing method of Australia La Woer chrysanthemum young shoot extract of tissue cultures characterized by comprising
(a) by Australia La Woer chrysanthemum young shoot of tissue cultures ethyl alcohol and 1,3 butylene glycol mixed solvent or ethyl alcohol and third
The step of in the mixed solvent of glycol impregnates 3~30 days and forms dipping solution；And
(b) the step of filtering the stain solution and obtaining Australia La Woer chrysanthemum young shoot extract of tissue cultures.
6. the manufacturing method of Australia La Woer chrysanthemum young shoot extract of tissue cultures according to claim 5, feature exist
In the weight of Australia La Woer chrysanthemum young shoot relative to the tissue cultures, the mixed solvent has 10~20 times of weight
Than.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR10-2014-0083018 | 2014-07-03 | ||
| KR1020140083018A KR101618180B1 (en) | 2014-07-03 | 2014-07-03 | Cosmetic composition comprising an extract of the tissue-cultured raoulia australis shoot |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105310923A CN105310923A (en) | 2016-02-10 |
| CN105310923B true CN105310923B (en) | 2018-12-25 |
Family
ID=55172500
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510232464.XA Active CN105310923B (en) | 2014-07-03 | 2015-05-08 | The manufacturing method of Australia La Woer chrysanthemum young shoot extract of cosmetic composition and tissue cultures |
Country Status (2)
| Country | Link |
|---|---|
| KR (1) | KR101618180B1 (en) |
| CN (1) | CN105310923B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR3061657B1 (en) * | 2017-01-09 | 2020-01-03 | Plant Advanced Technologies Pat | PREPARATION OF A ROOT EXTRACT OF FILIPENDULA ULMARIA AND COSMETIC USES |
| KR102420527B1 (en) * | 2017-09-22 | 2022-07-13 | 엘지이노텍 주식회사 | Camera module |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20020043840A (en) * | 2000-12-04 | 2002-06-12 | 서경배 | Method for stabilization of drug and decrease of skin-stimulation in the skin absorption having effect of removing wrinkle and protection wrinkle formation |
| EP2436757A2 (en) * | 2009-05-26 | 2012-04-04 | Unhwa Corporation | Plant stem cell derived from cambium of family asteraceae and method for the isolated culturing thereof |
-
2014
- 2014-07-03 KR KR1020140083018A patent/KR101618180B1/en active IP Right Grant
-
2015
- 2015-05-08 CN CN201510232464.XA patent/CN105310923B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20020043840A (en) * | 2000-12-04 | 2002-06-12 | 서경배 | Method for stabilization of drug and decrease of skin-stimulation in the skin absorption having effect of removing wrinkle and protection wrinkle formation |
| EP2436757A2 (en) * | 2009-05-26 | 2012-04-04 | Unhwa Corporation | Plant stem cell derived from cambium of family asteraceae and method for the isolated culturing thereof |
Non-Patent Citations (1)
| Title |
|---|
| Antiviral activity of raoulic acid from Raoulia australis against Picornaviruses;H.J. Choi et al;《Phytomedicine》;20091231(第16期);第35-39页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105310923A (en) | 2016-02-10 |
| KR20160004567A (en) | 2016-01-13 |
| KR101618180B1 (en) | 2016-05-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101677925B (en) | Compositions for improving skin conditions comprising alpha-bisabolol as an active ingrdient | |
| CN101641081B (en) | Compositions for improving skin conditions comprising matrine or its oxidized derivatives | |
| KR101990003B1 (en) | Manufacturing method of cosmetic composition containing Paeonia suffruticosa mixed extracts for anti-oxidation and whitening and cosmetic composition comprising thereof | |
| JP6054700B2 (en) | Desugaring agent and external preparation for skin | |
| KR102128974B1 (en) | Cosmetic composition for protecting skin from UV which comprises extract of bitter ground, persimmon leaf and ceramium kondoi | |
| CN103200929B (en) | The bioactive composition that comprises fig resin fraction and for reducing the method for the excessive appearance of cutaneous pigmentation | |
| KR101664025B1 (en) | Cosmetic composition containing natural herb extracts | |
| CN105310923B (en) | The manufacturing method of Australia La Woer chrysanthemum young shoot extract of cosmetic composition and tissue cultures | |
| KR101050613B1 (en) | Cosmetic composition containing plant extract having antioxidant effect and anti-wrinkle and moisturizing effect | |
| KR101489935B1 (en) | Method for preparing mugwort extract and cosmetic composition comprising mugwort extract prepared therefrom | |
| KR101518707B1 (en) | Cosmetic composition for skin whitening effect comprising hot water extract of strawberry plant powder as effective component | |
| KR20160000093A (en) | A cosmetic composition for antioxidizing and whitening containing the extracts of germinated crops | |
| KR101697235B1 (en) | Cosmetic Composition for Protection of Skin to Infrared Ray Comprising Hibiscus sabdariffa as Active Ingredient | |
| KR20130102712A (en) | Anti-heat aging effect of fermented limonia acidissima extract and its cosmetic usage to improve skin problems | |
| KR20180065998A (en) | Composition for skin-whitening comprising nanoparticle of callus derived from resveratrol-enriched rice | |
| KR101093901B1 (en) | A cosmetic composition comprising an extract of tissue cultured anoectochilus formosanus and a preparation method of the extract of tissue cultured anoectochilus formosanus | |
| KR20190049067A (en) | A cosmetic composition comprising extract of coffee silver skin and coffee grounds | |
| KR102176618B1 (en) | Composition comprising extract of zea mays linne | |
| KR101574248B1 (en) | Cosmetic composition comprising Reynoutria sachalinensis extracts for skin anti-wrinkle effect | |
| KR20130027227A (en) | Citrus flower compositions for improving skin conditions | |
| KR20120004021A (en) | Cosmetic composition comprising extracts of cultured wild ginseng root encapsulated in liposome and preparation method thereof | |
| KR20170007989A (en) | Whitening cosmetic composition including Chestnut shell extract, Albizziae Cortex extract and Kalopanax pictus extract | |
| KR20150010530A (en) | A cosmetic composition comprising critical extracts of pear | |
| KR20180137151A (en) | Cosmetic composition for preventing photo aging comprising Curcuma longa Linne leaf and Hibiscus manihot extracts | |
| KR102294241B1 (en) | Composition for antioxidant, whitening or anti-inflammation containing persimmon core extracts as an active ingredient |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |