CN109549913A - A kind of preparation method of the anti-ageing crease-resistant active principle extract of Chinese gall - Google Patents
A kind of preparation method of the anti-ageing crease-resistant active principle extract of Chinese gall Download PDFInfo
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- CN109549913A CN109549913A CN201710880599.6A CN201710880599A CN109549913A CN 109549913 A CN109549913 A CN 109549913A CN 201710880599 A CN201710880599 A CN 201710880599A CN 109549913 A CN109549913 A CN 109549913A
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- chinese gall
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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
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/987—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of species other than mammals or birds
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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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- Animal Behavior & Ethology (AREA)
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- Medicines Containing Plant Substances (AREA)
Abstract
The embodiment of the present invention provides a kind of preparation method of anti-ageing crease-resistant active principle extract of Chinese gall, it is related to field of cosmetic manufacture, being extracted using simple and quick method has skin antioxidant and anti-aging extract in Chinese gall, and the experiment proved that the active principle can significantly raise Collagen1, Elastin, Mn-SOD related gene, and it is remarkably improved T-SOD enzyme activity unit.The preparation method of the anti-ageing crease-resistant active principle extract of the Chinese gall includes: by Chinese gall crushed after being dried, the alcohol of addition 95% extracts, it extracts gained medicinal extract and petroleum ether degreasing is added, add ethyl acetate extraction effective component, gained ethyl acetate extracts medicinal extract, it is separated through macroporous absorbent resin, the anti-ageing crease-resistant active principle extract of Chinese gall is obtained after freeze-drying.
Description
Technical field
The present invention relates to a kind of systems of the anti-ageing crease-resistant active principle extract of field of cosmetic manufacture more particularly to Chinese gall
Preparation Method.
Background technique
Skin is maximum organ in human body, is the main protective barrier of body, the abnormal conditions of body can also be in skin
It is reflected on skin, therefore skin is most important to the health of body.Come along with accelerating to for aging society, research on anti-senescence
Increasingly deep, the skin of skin, especially exposure portion, which is that body aging is relatively early in the process, there is aging feature, and is to induce one
The aesthetic organ gazed at.The beautifying heat of the beautifying heat of world today's range, especially skin is deeply concerned.And the reason of back to nature
Thought makes people be more likely to selection natural plant extracts when selecting skin care item, therefore natural skin care item are deep by vast consumption
The favor of person.
Skin anti-aging is mainly focused on corium confluent monolayer cells, by RT-PCR and WB technology, from gene and protein level into
Row detection, as collagen, elastin, predominantly constitutes the collagenous fibres of skin corium using target gene and albumen as target
And elastic fibers, maintenance and support skin texture increase skin elasticity；Various anti-oxidant relevant enzymes, such as Mn-SOD, Cu/ in corium
Zn-SOD, CAT, GSH-Px, mainly by excessive free radical and peroxide in opposed body, to reduce its damage to skin
Wound increases the vigor of cell, improves skin aging.
As Chinese medicine and industrial chemicals, at home and abroad market enjoys high reputation Chinese gall.Chinese gall scientific name (Rhus
Chinensis Mill.) also known as hundred worm storehouses, fermented Chinese gall herb and tea leaves, gall, it is posted for the horned gall aphid or melaphis peitan Tsai et Tang female adult of Homoptera aphid section
It is born in Anacardiaceae plant " Chinese sumac " and its belongs to the tender leaf or petiole of other plant, stab and generate a kind of cryptomere consor object worm
Goitre, gained after baked drying.In addition, Chinese gall or a kind of medicinal material, can treat a variety of diseases.Chinese gall is grown in warm
Wet mountain area and hills is mainly distributed on six province such as Guizhou, Sichuan, Hubei, Hunan, Shaanxi, Yunnan in China main producing region, this
The Gallnut yield in a little areas accounts for about 90% or more of national total output.Gallic main component is that Chinese tannin is also known as five
Gall nut tannin has the function of that astringing lung-QI fall fire, relieving diarrhea with astringents, hemostasis, hygroscopic hold back wound.
Related Chinese gall composition Study, domestic and foreign literature concentrate on medicinal ingredient research aspect, medicinal ingredient baseset
In in the research and development of Chinese gall polysaccharide and tannin, so far, there has been no answer about Chinese gall polyphenol extract and correlation
For the exploitation of cosmetic material, the purpose of the present invention is to provide a kind of Chinese gall polyphenol applied to cosmetics is anti crease and anti senile
The preparation method of extract.And to its effective component carry out the crease-resistant research of anti-aging, human body patch experimental verification, specify its
Application in cosmetics.
Chinese patent application CN201210474797.X " Chinese gall anti-cancer effective component extract and its preparation method and use
The extracting method for relating to Chinese gall ethers extract on the way " is not directed to the extraction of Chinese gall polyphenol ingredient.
Chinese patent application CN201010579001.8 " a kind of gallnut polysaccharide composite and preparation method thereof ", is related to five
The extracting method of gall nut polysaccharide is not directed to Chinese gall polyphenol ingredient.
It is an object of the invention to provide a kind of crease-resistant active component of the anti-aging of nutgall extractive and its preparation sides
Method.
The second purpose of the invention is to provide application of the active component in preparation anti-aging anti-wrinkle cosmetic.
The third object of the present invention is to provide a kind of safe and reliable cosmetic material additive.
Summary of the invention
The embodiment of the invention provides a kind of preparation methods of the anti-ageing crease-resistant active principle extract of Chinese gall, using simple
Quick method, which is extracted, has skin antioxidant and anti-aging extract in Chinese gall, and the experiment proved that the active principle can
Significant up-regulation Collagen1, Elastin, Mn-SOD related gene, and it is remarkably improved T-SOD enzyme activity unit.
In order to achieve the above objectives, the embodiment of the present invention adopts the following technical scheme that
Chinese gall crushed after being dried is added 95% by a kind of preparation method of the anti-ageing crease-resistant active principle extract of Chinese gall
Alcohol extracts, and extracts gained medicinal extract and petroleum ether degreasing is added, and adds ethyl acetate extraction effective component, gained acetic acid second
Ester extracts medicinal extract, separates through macroporous absorbent resin, the anti-ageing crease-resistant active principle extract of Chinese gall is obtained after freeze-drying.
Further, the method specifically includes:
(1) it using Chinese gall as raw material, is crushed after Chinese gall impurity removing, passes through the worm corpse in screening removal Chinese gall, worm
The impurity such as excreta, purification material are kept in spare；
(2) alcohol is added and extracts 3-6h, extract in triplicate, filter, merging filtrate, 48-55 DEG C is concentrated under reduced pressure to obtain coarse extraction leaching
Cream is spare；
(3) coarse extraction medicinal extract adds appropriate amount of water at suspension, and the petroleum ether of 2-4 times of volume is added, and removes Chinese gall oil lipid component,
In triplicate；
(4) lower layer's water phase obtained by above-mentioned (3), concentration removal unrecovered oil ether, the ethyl acetate extraction for adding 2-4 times of volume are more
Phenol ingredient, in triplicate, Chinese gall polyphenol crude extract is concentrated under reduced pressure to obtain in combined ethyl acetate layer；
(5) Chinese gall polyphenol crude extract obtained by above-mentioned (4), is added suitable water and is dissolved, and carries out AB-8 macroporous absorption tree
Rouge separation is added deionized water elution and removes big polarity by Chinese gall polyphenol crude extract in resin material Static Adsorption 6-10h
Water soluble ingredient adds 70% alcohol and is eluted, and eluent is concentrated under reduced pressure to give the anti-ageing crease-resistant extract of Chinese gall；
(6) after the anti-ageing crease-resistant extract freeze-drying of Chinese gall obtained by above-mentioned (5) plus 50% pentanediol water carries out dissolving to obtain five times
The crease-resistant extracting solution of declining of son；
(7) the anti-ageing crease-resistant extracting solution of Chinese gall obtained by above-mentioned (6), through acute oral toxicity experiment, Multiple skin irritation test, urgency
Property Ocular irritation experiment, human body patch experimental verification be safety low-poison cosmetic material.
Further, it was 80 meshes that the Chinese gall in the step (1), which crushes dynamics,.
Further, the Extracting temperature in the step (2) is 55-65 DEG C；The alcohol concentration is 95%, institute in extraction
The solid-liquid ratio stated is 1:5-1:10.
Further, the applied sample amount in the step (5) and macroporous absorbent resin mass ratio 1:10-1:15；Gained elution
Volume is 3-4 times of column volume, flow velocity 3-5BV/h.
The pretreatment for the AB-8 macroporous absorbent resin that the present invention uses: with the immersion of 95% ethyl alcohol 24 hours or more, ethyl alcohol is used
Wet method dress post.Until continuing to be mixed into clarification with ethanol elution to eluent and water.Then pillar is eluted extremely with 5% dilute hydrochloric acid
7~8 column volumes are washed with water and take off until the pH value of eluent is neutral.And liquid level is put to resin and is flushed, so as to
Loading.
The gained anti-oxidant crease-resistant extracting solution of Chinese gall of the invention uses Folin-Phenol colorimetric method, using gallic acid as standard items,
Determine that the content of polyphenol substance and HPLC detection method analyze its main component retention time.Gained nutgall extractive is added
50% pentanediol water is dissolved, and can be applied to the additive of toner, lotion, essence, eye cream etc..
Beneficial effects of the present invention
(1) advantages of the present invention and effect are using solvent extraction, extraction and macroporous absorbent resin partition method, from Chinese gall
In extracted the active principle with anti crease and anti senile effect, which can significantly raise Collagen1, Elastin in vitro,
Mn-SOD related gene, and it is remarkably improved T-SOD enzyme activity unit.
(2) present invention has further expanded application of the nutgall extractive in skin care item and cosmetics.It can be used for out
Send out the skin care item such as skin activating water, re-surface intensive wrinkle correct, anti-wrinkle eye cream crease-resistant.
Detailed description of the invention
Attached drawing 1 is the HPLC spectrogram of nutgall extractive.HPLC spectrogram condition: mobile phase: 0.1% acetate methanol of A phase, D
0.1% acetic acid water of phase；0.01-20min A=10%；20-45min A=30%-100%；45-55min A=100%；C-
18,250mm*4.6mm, 5 μm；Flow velocity 1ml/min；Column oven: 30 DEG C；Sample volume: 10 μ l；Sample concentration 5mg/ml；Attached drawing 2
For gallic acid canonical plotting.Calibration curve equation y=79.264x+0.1515, R2=0.9978, range of linearity 2.5-12
μ g/ml brings normal equation into and calculates polyphenol concentration, according to calculation formula p=(c × v according to the light absorption value of measured sample
× n)/(w × 103) calculating polyphenol content；
Attached drawing 3 is influence of the nutgall extractive to SOD enzyme activity, and the anti crease and anti senile extracting solution of Chinese gall is remarkably improved SOD enzyme activity
Power, and effect is better than VC；
Attached drawing 4 is the variation of mRNA after nutgall extractive acts on for 24 hours.Chinese nutgall can significantly raise Collagen1,
Elastin, Mn-SOD related gene；
Attached drawing 5 is Chinese nutgall acute toxicity testing result.The anti-ageing crease-resistant extract of Chinese gall is low-toxicity product, IC50
Between 2500mg/kg and 5000mg/kg, configured Chinese nutgall is safe.
Attached drawing 6 is Chinese nutgall acute toxicity testing result.The anti-ageing crease-resistant extract of Chinese gall is low-toxicity product, is matched
The Chinese nutgall set is safe.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention is retouched in detail
It states.
Embodiment 1:
Chinese gall raw material 5kg is taken, is crushed, is sieved with 100 mesh sieve after removal of impurities, 95% alcohol 25L is added, is heated to 60 DEG C of leachings
It mentions, filters, recycle filtrate, in triplicate, filtrate is concentrated under reduced pressure in 48 DEG C, and concentrate is added after suitable quantity of water forms suspension and is added
5L petroleum ether extraction three times, removes liposoluble constituent；Lower layer's water phase adds the ethyl acetate extraction effective component of 5L, merges
Recycling design is concentrated under reduced pressure at -0.09MPa, 48 DEG C in extract liquor, obtains Chinese gall polyphenol coarse extraction medicinal extract, and -80 DEG C of medicinal extract cold
Dry Chinese gall polyphenol crude extract, total 1.8kg is lyophilized.40g crude extract is weighed, macroporous absorbent resin separation is carried out, collects 70%
Ethanol eluate, eluent are 4BV volume, are concentrated under reduced pressure, and freeze-drying obtains the anti-ageing crease-resistant extract of Chinese gall, through with forint
It is about 30.7% that phenol method, which measures polyphenol content, yield 24.5%, and HPLC detection spectrogram is shown in attached drawing 1.
Embodiment 2:
Chinese gall raw material 1kg is taken, is crushed after removal of impurities, is sieved, 95% alcohol 7L is added, 55 DEG C of extractions is heated to, filters,
Filtrate is recycled, in triplicate, filtrate is concentrated under reduced pressure in 50 DEG C, and concentrate is added after suitable quantity of water forms suspension and 2L petroleum ether is added
Extraction three times, removes liposoluble constituent；Lower layer's water phase add 2L ethyl acetate extraction effective component, combining extraction liquid, in-
0.09MPa, recycling design is concentrated under reduced pressure at 48 DEG C, obtains Chinese gall polyphenol coarse extraction medicinal extract, -80 DEG C of medicinal extract be freeze-dried five
The crease-resistant extract of gall nut, total 0.32kg.40g crude extract is weighed, macroporous absorbent resin separation is carried out, collects 70% ethanol elution
Liquid, eluent are 4BV volume, are concentrated under reduced pressure, and freeze-drying obtains Chinese gall polyphenol extract, through measuring polyphenol with Forint phenol method
Content is about 33.2%, yield 21.76%.
Embodiment 3:
Chinese gall raw material 0.5kg is taken, is crushed after removal of impurities, is sieved, 95% alcohol 3L is added, is heated to 58 DEG C of extractions, mistake
Filtrate is recycled in filter, and in triplicate, filtrate is concentrated under reduced pressure in 52 DEG C, and concentrate is added after suitable quantity of water forms suspension and 1.2L is added
Petroleum ether extraction three times, removes liposoluble constituent；Lower layer's water phase adds the ethyl acetate extraction effective component of 1.2L, merges extraction
Liquid is taken, recycling design is concentrated under reduced pressure at -0.09MPa, 48 DEG C, obtains Chinese gall polyphenol coarse extraction medicinal extract, -80 DEG C of medicinal extract freezings
Dry Chinese gall polyphenol crude extract, total 0.18kg.40g crude extract is weighed, macroporous absorbent resin separation is carried out, collects 70% second
Alcohol eluen, eluent are 4BV volume, are concentrated under reduced pressure, and freeze-drying obtains Chinese gall polyphenol extract, through being surveyed with Forint phenol method
Determining polyphenol content is about 31.8%, yield 23.4%.
Embodiment 4: anti-oxidant and activity of fighting against senium evaluation
The obtained anti-oxidant crease-resistant extract of Chinese gall determines the best use concentration by MTT as tested material using in example 1
25 μ g/ml survey anti-oxidant crease-resistant gene by gene water product examine, can significantly raise Collagen1, Elastin, Mn-SOD phase
Correlation gene, such as Fig. 3 and Fig. 5.
To the T-SOD enzyme activity detection after nutgall extractive effect, compareed as the result is shown with VC, Chinese nutgall can
T-SOD enzyme activity unit is significantly improved, as shown in Figure 4.
Table 1
Wherein, table 1 is Chinese nutgall Multiple skin irritation test result.The multiple skin thorn of Chinese nutgall as the result is shown
It is qualified to swash experiment.
Table 2
Wherein, table 2 is Chinese nutgall Ocular irritation experimental result.Chinese nutgall Ocular irritation is qualified as the result is shown.
Table 3
Wherein, table 3 is Chinese nutgall human body patch experimental result.Chinese nutgall human body patch experiment as the result is shown is closed
Lattice.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, any
Those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, and should all contain
Lid is within protection scope of the present invention.Therefore, protection scope of the present invention should be based on the protection scope of the described claims.
Claims (5)
1. a kind of preparation method of the anti-ageing crease-resistant active principle extract of Chinese gall, which is characterized in that by powder after Chinese gall drying
It is broken, 95% alcohol is added and extracts, extracts gained medicinal extract and petroleum ether degreasing is added, add ethyl acetate extraction effectively at
Point, gained ethyl acetate extracts medicinal extract, separates through macroporous absorbent resin, anti-ageing crease-resistant effective group of Chinese gall is obtained after freeze-drying
Divide extract.
2. the method according to claim 1, wherein the method specifically includes:
(1) it using Chinese gall as raw material, is crushed after Chinese gall impurity removing, passes through the worm corpse in screening removal Chinese gall, worm
The impurity such as excreta, purification material are kept in spare；
(2) alcohol is added and extracts 3-6h, extract in triplicate, filter, merging filtrate, 48-55 DEG C is concentrated under reduced pressure to obtain coarse extraction leaching
Cream is spare；
(3) coarse extraction medicinal extract adds appropriate amount of water at suspension, and the petroleum ether of 2-4 times of volume is added, and removes Chinese gall oil lipid component,
In triplicate；
(4) lower layer's water phase obtained by above-mentioned (3), concentration removal unrecovered oil ether, the ethyl acetate extraction for adding 2-4 times of volume are more
Phenol ingredient, in triplicate, Chinese gall polyphenol crude extract is concentrated under reduced pressure to obtain in combined ethyl acetate layer；
(5) Chinese gall polyphenol crude extract obtained by above-mentioned (4), is added suitable water and is dissolved, and carries out AB-8 macroporous absorption tree
Rouge separation is added deionized water elution and removes big polarity by Chinese gall polyphenol crude extract in resin material Static Adsorption 6-10h
Water soluble ingredient adds 70% alcohol and is eluted, and eluent is concentrated under reduced pressure to give the anti-ageing crease-resistant extract of Chinese gall；
(6) after the anti-ageing crease-resistant extract freeze-drying of Chinese gall obtained by above-mentioned (5) plus 50% pentanediol water carries out dissolving to obtain Chinese gall
The crease-resistant extracting solution of declining；
(7) the anti-ageing crease-resistant extracting solution of Chinese gall obtained by above-mentioned (6), through acute oral toxicity experiment, Multiple skin irritation test, urgency
Property Ocular irritation experiment, human body patch experimental verification be safety low-poison cosmetic material.
3. according to the method described in claim 2, it is characterized in that, it was 80 that the Chinese gall in the step (1), which crushes dynamics,
Mesh.
4. according to the method described in claim 2, it is characterized in that, the Extracting temperature in the step (2) is 55-65 DEG C；Institute
Stating alcohol concentration is 95%, and solid-liquid ratio described in extraction is 1:5-1:10.
5. according to the method described in claim 2, it is characterized in that, applied sample amount and macroporous absorbent resin in the step (5)
Mass ratio 1:10-1:15；Gained elution volume is 3-4 times of column volume, flow velocity 3-5BV/h.
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| CN102002114A (en) * | 2010-12-08 | 2011-04-06 | 张家界奥威科技有限公司 | Gallnut polysaccharide composite and preparation method thereof |
| CN102988421A (en) * | 2012-11-21 | 2013-03-27 | 江苏大学 | Extract of effective components of antineoplastic gallnut as well as preparation method and application thereof |
| CN103690567A (en) * | 2012-12-28 | 2014-04-02 | 中国医学科学院药用植物研究所 | Gallnut ethyl acetate extract as well as preparation and detection methods and application thereof |
| KR20170089093A (en) * | 2016-01-25 | 2017-08-03 | 주식회사 화수목 | A cosmetic composition for antioxidating comprising extracts of eisenia bycyclis, gallnut and rhubarb |
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2017
- 2017-09-26 CN CN201710880599.6A patent/CN109549913A/en active Pending
Patent Citations (5)
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| CN101433560A (en) * | 2007-11-13 | 2009-05-20 | 天津天士力制药股份有限公司 | Effective component of galla chinensis as well as preparation method and use thereof |
| CN102002114A (en) * | 2010-12-08 | 2011-04-06 | 张家界奥威科技有限公司 | Gallnut polysaccharide composite and preparation method thereof |
| CN102988421A (en) * | 2012-11-21 | 2013-03-27 | 江苏大学 | Extract of effective components of antineoplastic gallnut as well as preparation method and application thereof |
| CN103690567A (en) * | 2012-12-28 | 2014-04-02 | 中国医学科学院药用植物研究所 | Gallnut ethyl acetate extract as well as preparation and detection methods and application thereof |
| KR20170089093A (en) * | 2016-01-25 | 2017-08-03 | 주식회사 화수목 | A cosmetic composition for antioxidating comprising extracts of eisenia bycyclis, gallnut and rhubarb |
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| Title |
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Address after: 710042 room 112, floor 1, building A2, standard workshop community, Xi'an Textile Industrial Park, Baqiao District, Xi'an City, Shaanxi Province Applicant after: Shaanxi bairuiheng Health Technology Co.,Ltd. Address before: 523808 room 10, 1st floor, building 1, Dongguan Taiwan Biotechnology cooperation center, No.1 Taoyuan Road, Taiwan hi tech park, Songshanhu hi tech Industrial Development Zone, Dongguan City, Guangdong Province Applicant before: DONGGUAN NATURAL SALUTEM HEALTH TECHNOLOGY CO.,LTD. |
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Application publication date: 20190402 |