CN109303723A - Plum blossom embryo extract and its preparation method and application - Google Patents
Plum blossom embryo extract and its preparation method and application Download PDFInfo
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- CN109303723A CN109303723A CN201710623699.0A CN201710623699A CN109303723A CN 109303723 A CN109303723 A CN 109303723A CN 201710623699 A CN201710623699 A CN 201710623699A CN 109303723 A CN109303723 A CN 109303723A
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- plum blossom
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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Abstract
The invention belongs to field of plant extraction and daily chemicals field, specially a kind of plum blossom embryo extract is the full pulp that plum blossom spends embryo entirely;Plum blossom embryo extract has the function of promoting skin expression hyaluronic acid, type i collagen and elastin laminin, can be used as active constituent and be used to prepare skin preparations for extenal use, be conducive to skin and maintain higher moisture, keep skin wet, reduce the generation of wrinkle;And fibroblasts to secrete collagen, elastomer albumen and various kinds of cell reparative factor can be effectively facilitated, to repair the skin and delay skin aging of aging, has the function of anti-aging, there is moisturizing, anti-aging or repairs skin function.
Description
Technical field
The invention belongs to field of plant extraction and daily chemicals fields, and in particular to a kind of plum blossom embryo extract and its application.
Background technique
Skin is covered in entire human body surface, directly contacts with external environment, in addition to protect body from it is extraneous it is various because
It outside the injury of element, and is the mirror for reacting general health status.
Hyaluronic acid is a kind of acid mucopolysaccharide, also known as uronic acid, sodium hyaluronate;Hyaluronic acid has extraordinary water conservation
Effect, while being also a kind of multi-functional matrix, hyaluronic acid is distributed widely in partes corporis humani position.Wherein skin also contains largely
Hyaluronic acid.Human skin maturation and ageing process also with hyaluronic acid content and metabolism and change, it can be with
Improve skin-nourishing metabolism, make skin it is tender, it is smooth, go wrinkle, increase elasticity, delay senescence.
Collagen is the protein that content is most in the mammalian body, is the main component in extracellular matrix, widely distributed
In in the skin of human body, bone, muscle, cartilage, joint, hair tissue, play support, reparation, the triple of protection delay to decline
Old effect.Collagen is located in the skin corium of application on human skin to be synthesized by fibroblast, accounts for about the 95% of dermal components, wherein big portion
Being divided into type i collagen (90%), there are also small part type III collagens (10%);So type i collagen is main contained by dermal layer of the skin
Ingredient, with age, type i collagen is lost in skin, and atrophy, the aging phenomenons such as wrinkle occurs in content decline, skin.
Elastin laminin is the main component of elastomer.Elastin laminin is present in ligament, aorta, in skin and cartilage.
Although collagen can also need high resilience, therefore elastin laminin to extracellular matrix with intensity and toughness in skin
Take on important role in terms of maintaining elasticity of skin.Elastin laminin has the ability of stretching, extension and flatung by skin, is responsible for maintenance
With the elasticity of support skin.Elastin laminin decides the elasticity and flexibility of skin, to skin caused by the chemical factors such as light
Physiological aging process, having prevents aging and the regenerated effect of promotion.With age, elastin laminin in skin
Content is reduced, and is jointly reduced to network support required for compact skin is maintained, skin starts relaxation, has the agings such as microgroove
Phenomenon.
Therefore, how research increases the water content and enhancing elasticity of skin, for protecting health, the delaying skin of skin
Aging is of great significance.The synthesis for improving intracellular hyaluronic acid contents, promoting type i collagen and elastin laminin, can play
The effect of anti-aging.
Plum blossom is the bud of rosaceae cherry plum (Prunusmume Sieb.et Zucc.), and Japanese apricot opens for early spring
Colored defoliation small arbor, the REGIONALIZATION OF THE HENGDUAN MOUNTAINOUS REGION and the Yunnan-Guizhou Plateau that distribution center has a common boundary in China Sichuan, Yunnan and Tibet.Plum blossom is planted
Training is easy, and the service life is long;Extremely sensitive to temperature, florescence maximum affected by environment is hardy plant, can open at 0 DEG C or so
Flower, in the early time at florescence, when 4 DEG C of mean of dekan temperature or so, so that it may enter full-bloom stage.
Plum blossom is traditional famous flower in China, there is the title of the queen of flowers, has pleasant fragrance, the spiritual character to defy severe cold
Always by the high praise of the men of literature and writing and highly praise, and pine, bamboo and claim " three durable plants of winter: pine, bamboo and plum blossom ", and is " four gentleman " (plum, orchid, bamboo, chrysanthemum)
One of, it is very popular, have more than 3000 years cultivation histories, be now People's Republic of China's candidate's national flower it
One.
Plum blossom is widely used, and in addition to for watching, dried flower, leaf, root, kernel of plum blossom etc. can be used as medicine, and fresh flower can Titian
Essence, some plum blossom kinds can flowers and fruits dual-purpose, tied plum can processed goods, the smoked plum cream such as dried plum processed, green plum, preserved plum, dark plum be even more
Antibacterial, the sterilization good medicine for preventing and treating enteric infectious disease.
The main active of plum blossom has flavonoids and organic acid compound.The Huang of identification is separated from plum blossom at present
Ketones component has 17 kinds, including flavone aglycone (such as Quercetin, Isorhamnetin), oligosaccharides glycosides (e.g., rutin, Hyperoside, different Mongolian oak
Skin glycosides) and polysaccharide glycosides and its derivative;Organic acid ingredient in plum blossom is mainly chlorogenic acid substance.Kyoto, Japan medicine section
University carries out experiment discovery to experimental mouse with the certain ingredients of plum blossom, and flos mume general flavone extracting solution has suppression to Mushroom Tyrosinase
System activity mitigates cutaneous pigmentation to interfere the biosynthesis of melanocyte, improves the colour of skin.Research shows that flos mume methanol mentions
Object and its ethyl acetate layer, n-butanol layer, aqueous fraction is taken to show stronger DPPH free radical and O2—: free radical is clear
Except activity.
There is presently no Flos Mume extract is used to improve, delay skin aging, maintain the report of skin function, also do not have
It is found other methods and obtains active constituent from plum blossom, the report for delay skin aging.
Summary of the invention
The present invention is intended to provide a kind of plum blossom embryo extract and preparation method.
The present invention also by above-mentioned plum blossom embryo extract be applied to skin preparations for extenal use, especially have anti-aging, moisturizing or
Repair the cosmetics of the functions such as skin.
Technical solution are as follows: a kind of plum blossom embryo extract is the full pulp of plum blossom embryo.
Preferably, the plum blossom embryo is in bud to bud stage.It is furthermore preferred that plum blossom embryo is in bud stage.
Preferably, the plum blossom embryo is the flower embryo of armeniaca mume.
Above-mentioned plum blossom embryo extract can be prepared by the following method: take fresh plum blossom embryo (entirely spend on embryo) squeezing and/
Or mashing, obtain full pulp.Preferably, freezing squeezing and/or mashing are carried out at -18~-200 DEG C.
Preferably, fresh plum blossom embryo is refrigerated to -18~-200 DEG C after picking in 24 hours.
Preferably, freezing squeezing obtains full pulp at -18~-200 DEG C.More preferably, the temperature of squeezing is freezed
It is -50~-200 DEG C.In a preferred embodiment of the present invention, the temperature for freezing squeezing is -150~-200 DEG C.
Plum blossom embryo extract of the invention, does not generate toxicity to Skin Cell, can be used as functional additive and be used for
Prepare skin preparations for extenal use.
Plum blossom embryo extract can promote hyaluronic acid synthesis, improve intracellular hyaluronic acid and contain as active constituent
Amount promotes type i collagen synthesis and promotes the synthesis of elastin laminin, can be used as moisturizing, anti-aging or the activity for repairing skin function
Ingredient or functional additive are used for skin preparations for extenal use, realize function of delaying senility.Plum blossom embryo extract can also significantly improve
The hyaluronic acid contents of skin fibroblasts, help to maintain skin wet, have moistening effect, reduce and generate wrinkle;Also
It can effectively improve the expression quantity of type i collagen synthetic quantity and elastin laminin, so that elasticity of skin is maintained, delay skin aging.
Skin preparations for extenal use using above-mentioned plum blossom embryo extract as active constituent has moisturizing, anti-aging or reparation
The function of skin.
Skin preparations for extenal use using plum blossom embryo extract of the invention as active constituent, such as skin care item or cosmetics,
Have the function of to promote hyaluronic acid, I-type collagen or elastin laminin synthesis etc..
Skin preparations for extenal use can be toner, Essence, lotion, creams etc..
Hyaluronic acid is a kind of acid viscous polysaccharide, is distributed widely in partes corporis humani position, also contains in skin a large amount of saturating
Bright matter acid, has extraordinary water retention.Collagen is the main component in extracellular matrix, in human skin cell
It is synthesized by fibroblast, plays support, maintenance and protection.Elastin laminin is the main component of elastomer, is responsible for
Maintain and support the elasticity of skin.The moisture-keeping function of hyaluronic acid is conducive to skin and keeps moisture, can reduce the generation of dry line;I
The raising of Collagen Type VI and elastin laminin expression is able to maintain that elasticity of skin, further reaches anti-aging function.
After being handled with 0.03wt% plum blossom embryo extract, hyaluronic acid contents, type i collagen in fibroblast are closed
It is dramatically increased at the expression quantity of amount and elastin laminin, so that demonstrating plum blossom embryo extract is able to maintain that elasticity of skin,
Delay skin aging.
Additive amount of the plum blossom embryo extract in skin preparations for extenal use is 0.0001wt%~20wt%, preferably
0.001wt%~2wt%, more preferably 0.005wt%~0.5wt%.
Have present invention firstly provides plum blossom embryo extract and its compound and improves intracellular hyaluronic acid contents, rush
The effect synthesized into type i collagen and elastin laminin, suitable for the cosmetics with function of delaying senility.The preparation of extract
Method is simple, no solvent residue, can guarantee that its effective component is not lost, anti-oxidant and anti-aging effects are significant.
Detailed description of the invention
Fig. 1 is the experimental result that 2 plum blossom embryo extract of embodiment influences cell activity
Fig. 2 is the experimental result that 3 plum blossom embryo extract of embodiment promotes hyaluronic acid synthesis
Fig. 3 is the experimental result that 4 plum blossom embryo extract of embodiment promotes I-type collagen synthesis
Fig. 4 is the experimental result that 5 plum blossom embryo extract of embodiment promotes elastin laminin synthesis
Specific embodiment
Embodiment 1
The flower embryo that armeniaca mume is in the high activity stage is chosen, that is, is in the flower of bud stage, carries out artificial picked by hand;It adopts
By its quick freeze to -28 degree in 24 hours after plucking;Fresh full fruit is obtained using freezing Technology in Pressing under -196 degree
Slurry, as plum blossom embryo extract.
The whole low temperature of chilled quick-frozen and extraction process carries out, and full flower embryo is squeezed crushing and homogenate at low temperature, and without molten
Agent residual, to guarantee that its fresh and alive energy ingredient is lockable non-inactivation entirely, performance is more stable.
2 cell toxicity test of embodiment
Plum blossom embryo extract acquired in Example 1 is added distilled water or ultrapure water dilution, is used for subsequent experimental.
Cell culture: people's primary fibroblast, with 3000 cells/well kinds in 96 orifice plates, 37 DEG C, 5%CO2Item
With complete medium (DMEM 90%, FBS10%, dual anti-P/S 1%) under part, cultivated 48 hours in cell incubator.
Cell toxicity test step are as follows: plum blossom embryo extract (final concentration is added in the cell by culture
After 0.03wt%) co-culturing 24 hours, culture medium is removed, MTT 0.5mg/mL is added, supernatant is carefully sucked after 3 hours, is added
DMSO is detected under 550nm;Sample treatment group is not added as control, definition sample well is T, and untreated hole is NT,
It is considered as no cytotoxicity with ratio (T/NT) >=90% of T and NT, ratio < 90%, which is considered as, cytotoxicity.
In cell toxicity test, if the upgrowth situation for being added to extract is good, cell does not occur obvious dead
It dies.It then can tentatively judge that plum blossom embryo extract does not have toxicity to cell, can carry out follow-up test.
As a result such as Fig. 1, plum blossom embryo extract will not cause cytotoxicity to cell under 0.03wt% concentration.
The measurement of next step hyaluronic acid contents, Contents of collagen and elastin laminin is carried out with the concentration of 0.03wt% to close
At test.If hyaluronic acid contents are significantly raised relative to the groups of cells for being not added with extract, collagen content is increased, bullet
Property albumen synthesis also increase, deducibility plum blossom embryo extract be conducive to structure relevant to aging consolidate, finally play
Facilitate the effect of delay skin aging.
Embodiment 3 promotes hyaluronic acid synthesis
Cell culture processes such as embodiment 2.
After plum blossom embryo extract (final concentration 0.03wt%) is added in the cell of culture and co-cultures 24 hours, go
Supernatant is added not serum-containing media (DMEM 100%, dual anti-P/S 1%) and continues culture 24 hours, taken in orifice plate later
Clearly, it is operated according to Hyaluronan ELISA KIT specification, carries out the measurement of hyaluronic acid contents, detected under 450nm,
Standard curve is done with hyaluronic acid standard items (30~0ng/ml) series.Definition sample well is T, and untreated hole is NT.
As the result is shown as shown in Fig. 2, plum blossom embryo extract is in concentration 0.03%, hyaluronic acid contents are dramatically increased
To for 123.43% (promote delay skin aging effect of hyaluronic acid synthesis relative to untreated 100%),
Embodiment 4 promotes type i collagen synthesis
After plum blossom embryo extract (final concentration 0.03wt%) is added in the cell of culture and co-cultures 24 hours, go
Supernatant, not serum-containing media (DMEM 100%, dual anti-P/S 1%), which is added, is enriched with collagen 48 hours, takes in orifice plate later
Supernatant is operated according to COL-I ELISA KIT specification, is carried out the measurement of people's type i collagen content, is detected under 450nm, with
People's type i collagen standard items (20~0ng/ml) series does standard curve.Come with the amount of DNA that PicoGreen DNA kit measures
Demarcate the cell number in every hole.
As a result such as Fig. 3, from the figure 3, it may be seen that plum blossom embryo extract is when concentration is 0.03%, collagen content is than untreated
Kong Gao illustrates that the plum blossom embryo extract being added has the function of promoting type i collagen synthesis, and dramatically increases that type i collagen contains
Amount is 127.35% (relative to untreated hole 100%).
5 elastin laminin elastin fluorescence immunoassay expression analysis of embodiment
People's primary fibroblast is inoculated on 12 orifice plate cell climbing sheets with 5000/ hole, is cultivated 96 hours, hungry later
Then culture 6 hours with addition and is added without active matter (plum blossom embryo extract final concentration 0.03wt%) and is cultivated respectively
24 hours, the immunofluorescence dyeing that cell is Elastin was fixed later.
The cell climbing sheet for being added and not being added extract is taken respectively, sops up the culture medium on creep plate, and PBS is cleaned 2 times;It is cold
4 DEG C of fixations of methanol, PBS are cleaned 2 times;It Triton0.5% permeable membrane 15min, PBS cleaning 2 times, is closed later with 1%BSA/PBS,
1 that Elastin is added is anti-, and 4 stay overnight;Corresponding secondary antibody is added in next day and DAPI room temperature is protected from light and is incubated for 1h.Dyeing, is eventually adding
Mountant mounting, fluorescence microscopy microscopic observation are taken pictures.
As a result such as Fig. 4, and can be seen that the sample sets that 0.03% plum blossom embryo extract is added have by immunofluorescence analysis
The trend for increasing Elastin expression improves 24% than untreated Elastin expression, this illustrates that extract can be improved very
The expression of important feature albumen elastin laminin elastin, facilitates delay skin aging in skin.
Embodiment 6
The plum blossom embryo extract of embodiment 1 can be used as the active constituent flower embryo myogenic quintessence of skin preparations for extenal use, can
Remarkably promote hyaluronic acid, collagen especially I-type collagen, elastin laminin expression and synthesis.Flower embryo myogenic quintessence is made
It is added in skin preparations for extenal use such as cosmetics or skin care item for effective component or active constituent, cosmetics obtained or skin care
Product have moisturizing, crease-resistant smoothing wrinkle, anti-aging or delay skin aging, increase skin elasticity and repair skin function and other effects.
Dispersing agent, stabilizer, emulsifier, thickener etc. can also be added by plum blossom embryo extract and colored embryo myogenic is made
Quintessence active additive, such as palmityl hexapeptide -12 is added, finally the concentration used in cosmetics is 0.3%.
Conclusion: the experimental result of embodiment 2~5 is indicated using means ± CV, with Microsoft Excel statistical software
It is analyzed and processed.Compare two-by-two and examined using t, P < 0.05 * indicates statistical difference.Image analysis uses Image-Pro
Plus 7.0 carries out processing analysis, and the resultful data of institute are compared with untreated, and using untreated as 100%,
At least three Duplicate Samples (n=3) under the conditions of every kind.
One, cytotoxicity MTT experiment is shown, plum blossom embryo extract does not have cytotoxicity in concentration 0.03%.
Two, the effect of plum blossom embryo extract is in terms of delay skin aging
In order to verify the effect of plum blossom embryo extract is in terms of delay skin aging, we are from saturating after sample treatment
Three bright matter acid content, collage synthesis amount and elastin laminin expression quantity aspects are analyzed.If adding plum blossom embryonic complex object
Afterwards, cell growth state is good, by measuring hyaluronic acid contents, the synthetic quantity of collagen content and elastin laminin, if relatively
Rise in the control group content for being not added with plum blossom embryonic complex object, plum blossom embryonic complex object can be reflected for maintaining skin
Elasticity has effect, further reaches anti-aging function.
Plum blossom embryo extract concentration be 0.03% when, hyaluronic acid contents measurement, people's I Collagen Type VI assay and
The expression experiment display of elastin laminin Elastin immunofluorescence, hyaluronic acid, people's type i collagen content are all significantly increased.
1. adding plum blossom embryo extract (0.03wt%) has the delay skin aging function for promoting hyaluronic acid synthesis
Effect, hyaluronic acid contents are increased to 123.43%, increase 23.43% relative to untreated (100%).
2. the content of type i collagen dramatically increases after plum blossom embryo extract (0.03wt%) processing, content is untreated hole
127.35%, improve 27.35%.
3. immunofluorescence analysis has increase bullet the results show that the sample sets of plum blossom embryo extract (0.03wt%) are added
Property albumen Elastin expression trend;After the processing of plum blossom embryo extract, important feature albumen elastin laminin is expressed in corium
It is significantly increased, improves 24% compared with the control group.
It can be seen that plum blossom embryo extract, which is added, can promote the synthesis of hyaluronic acid, there is moisturizing and delaying skin
The effect of aging;Can promote the synthesis of type i collagen, improve the expression quantity of important feature elastin laminin in corium, be conducive to
Consolidating for the relevant structure of aging, facilitates delay skin aging.
Claims (10)
1. a kind of plum blossom embryo extract, which is characterized in that be the full pulp of plum blossom embryo.
2. plum blossom embryo extract described in claim 1, which is characterized in that the plum blossom embryo is the flower embryo of armeniaca mume.
3. plum blossom embryo extract described in claim 1, which is characterized in that the plum blossom embryo is in bud to bud stage.
4. the preparation method of any one of claims 1 to 3 plum blossom embryo extract, which is characterized in that take fresh plum blossom
The squeezing of flower embryo and/or mashing, obtain full pulp.
5. application of the described in any item plum blossom embryo extracts of claims 1 to 3 in terms of preparing skin preparations for extenal use.
6. application described in claim 5, which is characterized in that the plum blossom embryo extract as moisturizing, anti-aging or is repaired
The active constituent of multiple skin function.
7. application described in claim 5, which is characterized in that the plum blossom embryo extract is closed as promotion hyaluronic acid
At, promote I-type collagen synthesis or promote elastin laminin synthesis active constituent.
8. a kind of skin preparations for extenal use, which is characterized in that contain the described in any item plum blossom embryo extracts of claims 1 to 3.
9. a kind of compound of embryo extract containing plum blossom, which is characterized in that contain the described in any item plums of claims 1 to 3
Colored embryo extract is spent, at least one of dispersing agent, emulsifier, stabilizer, moisturizer, activity of fighting against senium ingredient are also contained.
10. application of the compound of the embryo extract containing plum blossom described in claim 9 in terms of preparing skin preparations for extenal use.
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