CN105250168B - A kind of high mountainous tea proferment pulp cosmetic and the preparation method and application thereof - Google Patents
A kind of high mountainous tea proferment pulp cosmetic and the preparation method and application thereof Download PDFInfo
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- CN105250168B CN105250168B CN201510639317.4A CN201510639317A CN105250168B CN 105250168 B CN105250168 B CN 105250168B CN 201510639317 A CN201510639317 A CN 201510639317A CN 105250168 B CN105250168 B CN 105250168B
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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
- 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
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
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Abstract
The invention discloses a kind of high mountainous tea proferment pulp cosmetic and its preparations.The high mountainous tea fermentation magma is prepared by following methods: high mountainous tea dry powder, water and bacterium solution being mixed to get initial system, fermented and cultured is carried out to the initial system, obtains high mountainous tea proferment pulp cosmetic.The zymophyte is saccharomyces cerevisiae.Application of the high mountainous tea proferment pulp cosmetic in the product that preparation has whitening and/or anti-senescence function also belongs to protection scope of the present invention.The product is cosmetics, concretely any one of facial mask, Essence or toner.High mountainous tea proferment pulp cosmetic prepared by the present invention is free of any chemical component, can use directly as the finished product of facial mask or Essence or toner, more natural than existing other products on the market, and any negative interaction will not be caused to skin；And the small active constituent of molecular weight can be obtained, it is easier to be fully absorbed by skin.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to a kind of high mountainous tea proferment pulp cosmetic and preparation method thereof with
Using.
Background technique
High mountainous tea refers to the produced tea between 1000 meters to 2600 meters of height above sea level, it is a kind of semi-fermented tea, belongs to oolong
Tea.Appearance is spherical or hemispherical.Contain many kinds of substance such as tea polyphenols more more than general tealeaves and caffeine, tool in high mountainous tea
There is adjusting fat metabolism, anticancer, anti-radiation and antibacterial, resisting age of skin reduce damage of the ultraviolet radiation in daylight to skin
And other effects, arousing brain, raising spirit, diuresis recover from fatigue and other effects.
What existing high mountain tea active matter extraction was commonly used is supercritical ultrasonics technology: high mountainous tea dry product adds a certain concentration after crushing
Ethyl alcohol, ultrasound a period of time, take supernatant after centrifugation, continue ultrasonic in ethanol solution, merge filtrate twice.But the party
Method introduces organic matter during the extraction process, not only breaks up environment, and there are potential security risks during application.
Summary of the invention
An object of the present invention is to provide a kind of preparation method of high mountainous tea fermentation magma.
The preparation method of high mountainous tea provided by the present invention fermentation magma, include the following steps: by high mountainous tea dry powder, water and
Bacterium solution is mixed to get initial system, carries out fermented and cultured to the initial system, obtains high mountainous tea fermentation magma.
In above-mentioned preparation method, the high mountainous tea dry powder is the tealeaves dry powder of high mountainous tea.
The size of the high mountainous tea dry powder is 20-50 mesh, concretely 30 mesh.
The high mountainous tea concretely Taiwan Alishan high mountainous tea.
The bacterial concentration is 104-107CFU/mL。
The proportion of the bacterium solution, high mountainous tea dry powder and water are as follows: 6-14mL:9-21g:320g, concretely 7mL:13g:
320g。
In the bacterium solution, strain is saccharomycete, concretely Saccharomyces Cerevisiae in S accharomyces cerevisiae.
The pH value of the bacterium solution is 6.5-7.5.
35-42 DEG C of the temperature of the fermented and cultured, specially 37 DEG C；Time is 19-26h, specially 21h.
Above-mentioned preparation method further includes on successively being sterilized, be centrifuged to the fermentation liquid obtained after the fermented and cultured, being taken
Clear liquid obtains the step of high mountainous tea fermentation magma.
The condition of the sterilizing are as follows: sterilising temp be 110-125 DEG C, specially 115 DEG C, sterilization time 17-35min,
Specially 20min.
The condition of the centrifugation are as follows: centrifugal rotational speed 4500-6700r/min, specially 5300r/min, centrifugation time are
18-45min, specially 20min, centrifugation radius are 9-12cm, specially 9cm.
The preparation-obtained high mountainous tea fermentation magma of the present invention also belongs to protection scope of the present invention.
The preparation-obtained high mountainous tea fermentation magma of the present invention is in the cosmetics that preparation has following at least one functions
Application or directly as in the cosmetics with following at least one functions application also belonging to protection scope of the present invention:
1) whitening；
2) anti-aging；
3) free radical is removed；
4) inhibit tyrosinase.
In above-mentioned application, the cosmetics are facial mask, Essence or toner.
The present invention provides the preparation method and application of a kind of high mountainous tea fermentation magma, this method selects more suitable bacterium
Kind, keep product more natural, and do not need the foreign substances such as addition enzyme, not only saves production cost, but also maximumlly simplify
Production stage, makes the fermentation technique can be realized mass production, industrialized production, and can fully ensure that the steady of product quality
It is qualitative.Be experimentally confirmed: high mountainous tea fermentation magma prepared by the present invention, can be directly as facial mask without any chemical component
Or the finished product use of Essence or toner, it is more natural than existing other products on the market, it will not be caused to skin any
Negative interaction；And the small active constituent of molecular weight can be obtained, it is easier to that skin is allowed to fully absorb.
Detailed description of the invention
Fig. 1 is the active peptides molecular weight distribution HPLC map of high mountainous tea proferment pulp cosmetic.
Fig. 2 is the removing DPPH free radical result schematic diagram of high mountainous tea proferment pulp cosmetic.
Fig. 3 inhibits tyrosinase activity schematic diagram for high mountainous tea proferment pulp cosmetic.
Specific embodiment
The present invention will be described below by way of specific embodiments, but the present invention is not limited thereto.
Experimental method used in following embodiments is conventional method unless otherwise specified；Institute in following embodiments
Reagent, material etc., are commercially available unless otherwise specified.
Fluid nutrient medium in following embodiments is made of solute and solvent, and solvent is water, solute and its in Liquid Culture
Concentration in base are as follows: glucose 20g/L, peptone 20g/L, yeast extract 10g/L, pH value 7.0.
YPD culture medium in following embodiments is made of solute and solvent, and solvent is water, solute and its liquid medium within
In concentration are as follows: yeast extract 1% (mass fraction), tryptone 2% (mass fraction), glucose 2% (mass fraction),
PH value is 7.0.
High mountainous tea in following embodiments is Taiwan Alishan high mountainous tea.
The ATCC number of Saccharomyces Cerevisiae in S accharomyces cerevisiae in following embodiments is 26603, can be from
ATCC is obtained.
The preparation method of embodiment 1, a kind of high mountainous tea fermentation magma
1, the activation of strain
One ring of picking Saccharomyces Cerevisiae in S accharomyces cerevisiae bacterium colony is put into shaking table in fluid nutrient medium
By actication of culture, saccharomyces cerevisiae bacterium solution after being activated.
2, the purifying of strain
Bacterium solution gradient dilution bed board after the activation that step 1 is obtained, to obtain single colonie, ferment of being made wine after purification
It is female.
3, the expansion culture of strain
The strain after purification that step 2 is obtained is inoculated into YPD culture medium (pH value 7.0), in 25 DEG C of shaking table
Culture, as OD value=0.4-0.8, obtaining zymophyte bacterium solution, (strain is in logarithmic phase, concentration 104-107CFU/ml)。
4, the acquisition of high mountainous tea fermentation magma
The zymophyte bacterium solution that 7mL step 3 obtains is inoculated into (zymophyte bacterium in the high mountainous tea dry powder of 13g and the water of 320g
Liquid concentration is 104-107CFU/ml), obtain fermentation system (strain total amount and the proportion of high mountainous tea are 7mL:13g in fermentation system:
320g)；Fermentation system is fermented 21 hours in 37 DEG C of shaking tables, obtains tunning；By 115 DEG C of high pressure sterilizations of tunning
20min inactivates bacterium, the tunning after being sterilized；It is in 5300r/min, centrifugation radius by the tunning after sterilizing
It is centrifuged 20min under conditions of 9cm, abandons precipitating, collects supernatant, as high mountainous tea fermentation magma.
The application of embodiment 2, high mountainous tea proferment pulp cosmetic:
One, the property of high mountainous tea proferment pulp cosmetic:
The embodiment prepares that the appearance of resulting high mountainous tea proferment pulp cosmetic (supernatant) is thick liquid, color is
It is faint yellow to brown color.PH value 5.2-6.3, viscosity 200-500cP, soluble solids content 1.5-5.0%, total plate count are small
In 50CFU/ml, no pathogenic bacteria detection.According to hygienic standards for cosmetics GB7916-87, cosmetics total number of bacteria is not higher than
1000CFU/ml, so this extractive from fermentative meets cosmetics quality requirement.
Constituent analysis is carried out to the high mountainous tea proferment pulp cosmetic, protein detection method is referring to GB5009.5-2010；
Thick many candies detection method is referring to GB/T 5009.8-2008；Flavones detection method is referring to GB/T 5009.124-2003；Total phenol inspection
For survey method referring to GB/T 8313-2008, acquired results are as follows:
High mountainous tea proferment pulp cosmetic produced by the present invention 5.73g/kg containing protein, Thick many candies 7.04g/kg, total Huang
Ketone 0.17g/kg (in terms of rutin), total phenol 0.28g/kg.
Two, the safety detection of high mountainous tea proferment pulp cosmetic
Human body patch test is mainly used for detecting the irritation of cosmetic end product or raw material.The present invention is to embodiment 1
The high mountainous tea fermentation magma of acquisition carries out human closed formula patch test, it is intended to assess its potential skin irritation.
1, subjects
Suitable 30 people of volunteer is selected, the range of age was randomly choosed at 18-60 years old.
2, test method
If solid sample or semisolid sample, then weigh 0.020g-0.025g be placed in it is spare in spot tester；If liquid
Body sample then measures 0.025mL-0.2mL and is added dropwise on filter paper, then filter paper is placed in spot tester.After the present invention uses
Person, each sample are respectively provided with blank control, be added in control spot tester hole with the sample solvent of sample equivalent, such as distilled water or
Olive oil.
Experiment position is selected as human body back, and spot tester fixation is pasted on subject back using nonirritant adhesive tape.
Test period continues for 24 hours.It is accurate, credible and scientific for test result, dduring test volunteer as requested, Bu Nengzhai
Fall spot tester, recipient site can not also be made to contact water.It removes spot tester afterwards for 24 hours, after standing 30min, impression is waited to disappear, observation
The reaction of skin.If test result is feminine gender, need to observe again respectively with 48h for 24 hours after patch test primary.
3, test result
Patch test results are as shown in table 1:
"-"=negative reaction；
" ± "=suspicious reaction: only faint erythema；
"+"=weakly positive reaction (erythematous response): erythema, oedema, can have papule at infiltration；
" ++ "=strong positive reaction (bleb reaction)；Erythema, infiltration, oedema, papule, bleb；Reaction can exceed tested area；
" +++ "=extremely strong positive reaction (reaction of amalgamation bleb)；Obvious erythema, severe infiltration, oedema, amalgamation blister
Rash；Reaction exceeds tested area.
According to cosmetics health specification 2007, human body patch test judgment criteria are as follows: occur 1 grade of skin in 30 subjects
The number of adverse reaction is more than 5 or the number of second level skin adverse reaction is more than 2, or any 1 three-level or three-level occurs
When the above skin adverse reaction, then determine that tested material has adverse reaction to human body, has no adverse reaction to human body conversely, being then considered as.
As can be seen from Table 1: the high mountainous tea proferment pulp cosmetic that embodiment 1 obtains generates 2 suspicious reactions, explanation
High mountainous tea proferment pulp cosmetic provided by the invention all has higher safety.
The patch test results for the high mountainous tea proferment pulp cosmetic that table 1, embodiment 1 obtain
Three, the molecular size range detection of high mountainous tea proferment pulp cosmetic
Molecular size range is to check whether an important factor for being able to carry out Cutaneous permeation.The present invention passes through high performance liquid chromatography
(HPLC) molecular size range of high mountainous tea fermentation magma prepared by measurement embodiment 1.Specific step is as follows:
1, the preparation of the reagent and solution tested
(1) mobile phase: acetonitrile: water: trifluoroacetic acid=30:70:0.1 (v/v/v).
(2) sample preparation: being the sample of 5mg/ml by solvent compound concentration of mobile phase, then with microporous barrier (0.45 μm)
For sample introduction after filtering.
(3) prepared by standard sample: by bovine serum albumin (Mr 67000), B12 (Mr 1335), oxidized form of glutathione (Mr
614) it is made into mixed mark, the concentration of every kind of substance is 5mg/ml.
2, test method
By the standard items of three kinds of known molecular amounts: bovine serum albumin(BSA) (Mw67000), vitamin B12(Mw1335) it and aoxidizes
Type glutathione (Mw614) passes through high performance liquid chromatograph (Waters company) reference literature " protease hydrolyzed wheat such as Wang Chengzhong
Method in the research grain processing that polypeptide molecular weight is distributed in embryo egg bletilla enzymolysis liquid, 2012:37 (2) " carries out HPLC points
The standard curve of molecular weight logarithm and elution time is drawn in analysis.According to gel column permeation chromatography principle, the big object of polypeptide molecular weight
Matter first is eluted out, and the logarithm of elution time and molecular weight is in a linear relationship.It will be in embodiment 1 according still further to same method
High mountainous tea fermentation magma prepared by experimental group 1, experimental group 2 and experimental group 3 carries out HPLC analysis, when obtaining the elution of respective peaks
Between, elution time is substituted into standard curve, the molecular size range of the high mountainous tea fermentation magma of the preparation of embodiment 1 can be obtained.
Chromatographic condition: chromatographic column: TsK gel 2000SWXL 300mm × 7.8mm；Detection wavelength: UV 280nm；Flow velocity:
1ml/min；Column temperature: 30 DEG C.
3, test result
The molecular weight of standard substance and elution time it is as shown in table 2, figure is done according to the logarithm of molecular weight and elution time
Obtain standard curve: y=-2.4738x+17.656, R2=0.9998.Wherein y is elution time (min)；X is molecular weight pair
Number.
Table 2, the molecular weight of standard substance and elution time
The HPLC map of high mountainous tea fermentation magma prepared by experimental group 1 is as shown in Figure 1: it can be seen from the figure that high mountainous tea
It mainly include 1 component peaks in the map for magma of fermenting, elution time is substituted into standard curve: y by elution time 11.58min
=-2.4738x+17.656, the molecular weight for calculating appearance is 285.59Da, and according to calculated by peak area high mountainous tea fermentation magma
Account for 99.11%.It is generally acknowledged that molecular weight 500Da or so activity peptide components are more easily absorbed by the skin, so in high mountain tea fermented product
In the presence of a large amount of absorbable active peptides.
Four, the antioxygenic property detection of high mountainous tea proferment pulp cosmetic
DPPH is a kind of organic free radical of early stage synthesis, is commonly used to the hydrogen supply capacity of assessment polyphenoils, it is organic
It is highly stable in solvent, it is purple, and have a characteristic absorption peak, when encountering free radical scavenger, the orphan of DPPH at place
Electronics is paired and it is made to fade, that is, is become smaller in the light absorption value of maximum absorption wave strong point.It therefore, can be by measuring extinction
The variation of value is to evaluate sample to the elimination effect of DPPH free radical.
The specific experiment step of DPPH radicals scavenging experiment are as follows:
(1) prepare liquid and 2 × 10 of isometric (generally 3mL) is taken-4The DPPH solution of mol/L mixes (A1Pipe)；
(2) isometric dehydrated alcohol (determinand solvent) and 2 × 10 is taken-4The DPPH solution of mol/L mixes (A2Pipe)；
(3) isometric dehydrated alcohol and prepare liquid is taken to mix (A3Pipe)；
(4) after reacting 30min, A is surveyed at 517nm1、A2、A3Pipe absorbance value.
Clearance rate calculation formula are as follows: clearance rate (%)=[(A2+A3)-A1]/A2
High mountainous tea proferment pulp cosmetic is made to DPPH radicals scavenging effect curves, sees attached drawing 2.
As shown in Figure 2, IC of the 1 gained high mountainous tea proferment pulp cosmetic of embodiment to DPPH scavenging effect50=1.18%,
Illustrate that there is high mountainous tea proferment pulp cosmetic stronger oxidation resistance can remove free radical, promote cell metabolism, enhancing is thin
Born of the same parents' vigor improves the structure and function of body, improves body vitality to delay cell senescence and plays the work of its anti-aging
With.
Five, the white-skinned face function analysis of high mountainous tea proferment pulp cosmetic
Tyrosinase is the key enzyme of melanogenesis, it controls the forming process of melanocyte, and level of activity is to pigment
Deposition plays a major role.Current market sales of many whitenings, nti-freckle product are all to inhibit tyrosinase to reach whitening and make
With, therefore be to evaluate the main indicator of whitening cosmetic to the power of tyrosinase inhibitory action.
The whitening function of sample is evaluated by measurement sample the influence of tyrosinase, and the specific method is as follows:
Solution is configured by table 3:
Table 3, solution prepare list
| Unit (mL) | C1 | C2 | T1 | T2 |
| L-tyrosine | 2 | 2 | 2 | 2 |
| Sample | 0 | 0 | 2 | 2 |
| PBS | 4 | 5 | 2 | 3 |
| Tyrosinase | 1 | 0 | 1 | 0 |
| Total volume | 7 | 7 | 7 | 7 |
Note: C1And T11mL tyrosinase, enzyme activity 100U/mL is added.
(1)C2Pipe prepares shake up after, return to zero under the heating water bath 10min in 37 DEG C of water-baths, wavelength 475nm.
(2)C1Pipe solution, which prepares, to be shaken up, and after 37 DEG C of water-bath 10min, is added tyrosinase 1ml, is continued water-bath 10 minutes, measurement
C1Absorbance value.
(3) (1) (2) same method is pressed, with T2Zeroing measurement T1Absorbance value.
(4) sample is calculated to the maximum inhibition T (%) of tyrosinase.T (%)=(C1-T1)/C1× 100%
Experiments have shown that high mountainous tea proferment pulp cosmetic is 57.69% to the inhibiting rate of tyrosinase activity, 1% black bearberry
Glycosides is shown in attached drawing 3 to the inhibiting rate 99.52% of tyrosinase activity.That is the whitening effect of high mountainous tea proferment pulp cosmetic is suitable
In the effect of 0.580% arbutin, there is certain white-skinned face function.
Claims (9)
1. a kind of preparation method of high mountainous tea fermentation magma, includes the following steps: for high mountainous tea dry powder, water and bacterium solution to be mixed to get
Initial system carries out fermented and cultured to the initial system, obtains high mountainous tea fermentation magma；
The bacterial concentration is 104-107CFU/mL；
The proportion of the bacterium solution, high mountainous tea dry powder and water are as follows: 6-14mL:9-21g:320g；
35-42 DEG C of the temperature of the fermented and cultured, time 19-26h；
In the bacterium solution, strain is saccharomyces cerevisiae (Saccharomyces cerevisiae).
2. preparation method according to claim 1, it is characterised in that: the high mountainous tea dry powder is that the tealeaves of high mountainous tea is dry
Powder.
3. preparation method according to claim 2, it is characterised in that: the high mountainous tea is Taiwan Alishan high mountainous tea.
4. preparation method according to any one of claim 1-3, it is characterised in that: the size of the high mountainous tea dry powder is
20-50 mesh.
5. preparation method according to any one of claim 1-3, it is characterised in that: the preparation method further includes to institute
State the step of gained fermentation liquid after fermented and cultured is successively sterilized, is centrifuged, supernatant is taken to obtain high mountainous tea fermentation magma.
6. preparation method according to claim 5, it is characterised in that: the condition of the sterilizing are as follows: sterilising temp 110-
125 DEG C, sterilization time 17-35min；
The condition of the centrifugation are as follows: centrifugal rotational speed 4500-6700r/min, centrifugation time 18-45min, centrifugation radius are 9-
12cm。
7. the high mountainous tea fermentation magma obtained by preparation method of any of claims 1-6.
8. application of the high mountainous tea fermentation magma as claimed in claim 7 in the cosmetics that preparation has following at least one functions
Or directly as the application in the cosmetics with following at least one functions:
1) whitening；
2) anti-aging；
3) free radical is removed；
4) inhibit tyrosinase.
9. application according to claim 8, it is characterised in that: the cosmetics are facial mask, Essence or toner.
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| CN105925617A (en) * | 2016-06-16 | 2016-09-07 | 北京工商大学 | Radix pueraiae fermentation protoplasm cosmetic product and preparation method and application thereof |
| CN107049912A (en) * | 2017-01-10 | 2017-08-18 | 韩后化妆品股份有限公司 | Lightening compositions and its application containing green tea fermentation filtrate |
| CN111616997A (en) * | 2020-06-05 | 2020-09-04 | 上海全丽生物科技有限公司 | Junshan honeysuckle tea fermentation raw stock and preparation method and application thereof |
| CN113827521A (en) * | 2021-10-18 | 2021-12-24 | 上海应用技术大学 | Preparation method of tea powder fermentation liquor and application of tea powder fermentation liquor in cosmetics |
| CN116549539A (en) * | 2022-01-30 | 2023-08-08 | 上海全丽生物科技有限公司 | Saffron fermentation product, external composition containing saffron fermentation product and having oil control and red dispelling effects, and application thereof |
| CN114989318A (en) * | 2022-05-09 | 2022-09-02 | 孙芙蓉 | Moisturizing and whitening polysaccharide and application thereof in preparing moisturizing and whitening cosmetics |
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| CN104257545A (en) * | 2014-09-18 | 2015-01-07 | 北京工商大学 | Rhodiola rosea fermentation protoplasm cosmetic and preparation method thereof |
| KR101485896B1 (en) * | 2014-10-27 | 2015-01-26 | 주식회사 더트루메틱 | Cosmetic composition for improving anti-oxidation, anti-inflammatory and atopic skin and method of preparing the same |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104257545A (en) * | 2014-09-18 | 2015-01-07 | 北京工商大学 | Rhodiola rosea fermentation protoplasm cosmetic and preparation method thereof |
| KR101485896B1 (en) * | 2014-10-27 | 2015-01-26 | 주식회사 더트루메틱 | Cosmetic composition for improving anti-oxidation, anti-inflammatory and atopic skin and method of preparing the same |
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