CN105213253A - A kind of compound recipe Synsepalum duleificum leaf extract and the application in skin-lightening cosmetic thereof - Google Patents
A kind of compound recipe Synsepalum duleificum leaf extract and the application in skin-lightening cosmetic thereof Download PDFInfo
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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
- 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
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
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Abstract
The invention discloses a kind of compound recipe Synsepalum duleificum leaf extract, it is prepared from herbal raw material, described herbal raw material is made up of major ingredient and adjuvant, described major ingredient is Synsepalum duleificum leaf, percentage by weight is 30% ~ 50%, adjuvant is the combination of any three kinds in Radix Paeoniae Rubra, Fructus Atriplicis Sibiricae, Olibanum, Rhizoma Curcumae Longae, the Radix Astragali, and percentage by weight is respectively 20% ~ 30%, 10% ~ 30% and 10% ~ 20%.The invention also discloses the preparation method of compound recipe Synsepalum duleificum leaf extract, comprise preparation herbal raw material, pretreatment, extraction, coarse filtration, fine straining, concentrate, redissolve and sterilizing.The invention discloses the application of described compound recipe Synsepalum duleificum leaf extract in skin-lightening cosmetic.Compound recipe Synsepalum duleificum leaf extract of the present invention has antioxidant activity and tyrosinase inhibitory activity concurrently, and compensate for the deficiency that Synsepalum duleificum leaf only has stronger antioxidant activity, flavones content is higher, and tyrosinase inhibition rate is also higher.
Description
Technical field
The present invention relates to cosmetic field, particularly relate to a kind of compound recipe Synsepalum duleificum leaf extract and the application in skin-lightening cosmetic thereof.
Background technology
Natural Synsepalum duleificum leaf, belongs to tropical evergreen shrub, evergreen all the year round, national treasure level rare species, embargo.Originate in West Africa, Ghana, the Congo one band, the sixties in 20th century, when Premier Zhou Enlai accesses to West Africa, Republic of Ghana gives premier Zhou using Synsepalum duleificum as state's gift.After this, start, in China's cultivation, to be rich in crude fibre, crude protein, crude fat, VC, polyphenol, polysaccharide, pyrite class material, is mainly used in health care of food product, though application has no report in cosmetic field, but because its polysaccharide, polyphenol, Flavonoid substances content are higher, there is in cosmetics using value potential greatly.Comparatively multi-nutrient is contained in Synsepalum duleificum leaf, as:
Synsepalum duleificum leaf nutritional labeling and containing scale
Summary of the invention
The technical problem to be solved in the present invention be to provide a kind of have antioxidant activity and tyrosinase inhibitory activity concurrently compound recipe Synsepalum duleificum leaf extract, preparation method and the application in skin-lightening cosmetic thereof.
A kind of compound recipe Synsepalum duleificum leaf extract, described compound recipe Synsepalum duleificum leaf extract is prepared from herbal raw material, described herbal raw material is made up of major ingredient and adjuvant, described major ingredient is Synsepalum duleificum leaf, percentage by weight is 30% ~ 50%, adjuvant is the combination of any three kinds in Radix Paeoniae Rubra, Fructus Atriplicis Sibiricae, Olibanum, Rhizoma Curcumae Longae, the Radix Astragali, and percentage by weight is respectively 20% ~ 30%, 10% ~ 30% and 10% ~ 20%.
Compound recipe Synsepalum duleificum leaf extract of the present invention, the preparation method of wherein said compound recipe Synsepalum duleificum leaf extract comprises the steps:
(1) described herbal raw material is prepared in proportion;
(2) pretreatment: carry out the broken process of ovendry power after being chosen by the foreign body in described herbal raw material, obtain herbal raw material powder, by four kinds of herbal powder mix homogeneously, makes Synsepalum duleificum leaf compound;
(3) extract: described Synsepalum duleificum leaf compound and solvent are mixed by weight 1:11 ~ 18, extracts 0.5 ~ 1 hour at the temperature of 26 ~ 45 DEG C, obtain extraction solution; Described solvent is pure water, the alcoholic solution of 40% ~ 95% or the acetone soln of 40% ~ 95%;
(4) coarse filtration: by described extraction solution coarse filtration, obtains compound recipe Synsepalum duleificum leaf extract coarse filtration liquid;
(5) fine straining: by described compound recipe Synsepalum duleificum leaf extract coarse filtration liquid fine straining, obtain compound recipe Synsepalum duleificum leaf extract fine straining liquid;
(6) concentrated: by described compound recipe Synsepalum duleificum leaf extract fine straining liquid, to obtain compound recipe Synsepalum duleificum leaf extract concentrated solution;
(7) redissolve: adopted by described compound recipe Synsepalum duleificum leaf extract concentrated solution the butanediol aqueous solution of 1:20 to redissolve, obtain compound recipe Synsepalum duleificum leaf extract redissolution thing;
(8) sterilizing, by the sterilizing of described compound recipe Synsepalum duleificum leaf extract redissolution thing, obtains compound recipe Synsepalum duleificum leaf extract.
Compound recipe Synsepalum duleificum leaf extract of the present invention, wherein, is extracted as stirring and refluxing and extracts or supersound extraction described in step (3).
The application of compound recipe Synsepalum duleificum leaf extract of the present invention in skin-lightening cosmetic.
Application of the present invention, wherein, in described skin-lightening cosmetic, described compound recipe Synsepalum duleificum leaf extract accounts for 0.2% ~ 8%.
Application of the present invention, wherein, one or more in solvent, emollient, solubilizing agent, antioxidant, pH adjusting agent, penetrating agent, emulsion stabilizer, antiseptic, wetting agent, thickening agent, nertralizer, chelating agen, surfactant, emulsifying agent, essence, coloring agent, sunscreen, skin conditioning agent, active matter and functional additive are also comprised in described skin-lightening cosmetic.
Application of the present invention, wherein, described skin-lightening cosmetic is astringent, facial film, facial cream, gel, emulsion, essence.
Compound recipe Synsepalum duleificum leaf extract of the present invention and the application difference from prior art in skin-lightening cosmetic thereof are:
Synsepalum duleificum leaf has good antioxidant activity, but there is no the function of restraint of tyrosinase activity, want to play its outstanding white-skinned face function, need consider to carry out compound recipe extraction, while giving full play to Synsepalum duleificum leaf antioxidant activity, carry more excellent skin-care effect.
About the selection of compound recipe, according to theory of Chinese medical science " dialectical, overall, comprehensive " and the monarch and his subjects' principles of formulating prescriptions " monarch, minister, help, make ", carry out composite, wherein determine that Synsepalum duleificum leaf is the main status of " monarch ", the Chinese herbal medicine selecting disposition with it to match carries out composite prescription, obtain flavone, polyphenols for basis for estimation to maximize, the optimal design of extraction process is carried out in centre, is applied in all kinds of State of cosmetics to reaching the best extracting solution of white-skinned face function.
Synsepalum duleificum leaf and Radix Paeoniae Rubra, Fructus Atriplicis Sibiricae, Olibanum, Rhizoma Curcumae Longae, the Radix Astragali etc. carry out composite by the present invention, have antioxidant activity and tyrosinase inhibitory activity concurrently, compensate for the deficiency that Synsepalum duleificum leaf only has stronger antioxidant activity, the present invention by Synsepalum duleificum leaf compound by pretreatment, heat of stirring reflux, extract, coarse filtration, fine straining, concentrated, redissolve, the process of sterilizing makes compound recipe Synsepalum duleificum leaf extract, its flavones content is higher, tyrosinase inhibition rate is also higher, can be used as white-skinned face function composition and is applied in cosmetics.
Detailed description of the invention
Embodiment 1
Herbal raw material is carried out pretreatment, makes herbal raw material powder;
Synsepalum duleificum leaf, Radix Paeoniae Rubra, Fructus Atriplicis Sibiricae, the Radix Astragali are made Synsepalum duleificum leaf compound according to the ratio of 30%, 30%, 20%, 20% respectively;
Synsepalum duleificum leaf compound and pure water are mixed by weight 1:11, makes mixture, this mixture is heated to about 26 DEG C, supersound extraction 30 minutes, make Synsepalum duleificum leaf compound and extract solution;
By Synsepalum duleificum leaf compound and extraction solution coarse filtration, make compound recipe Synsepalum duleificum leaf extract coarse filtration liquid;
By compound recipe Synsepalum duleificum leaf extract coarse filtration liquid fine straining, make compound recipe Synsepalum duleificum leaf extract fine straining liquid;
Compound recipe Synsepalum duleificum leaf extract fine straining liquid is concentrated, makes compound recipe Synsepalum duleificum leaf extract concentrated solution;
The butanediol aqueous solution of compound recipe Synsepalum duleificum leaf extract concentrated solution 1:20 is redissolved, makes compound recipe Synsepalum duleificum leaf extract redissolution thing;
By the sterilizing of compound recipe Synsepalum duleificum leaf extract redissolution thing, make compound recipe Synsepalum duleificum leaf extract.
Embodiment 2
Synsepalum duleificum leaf, Radix Paeoniae Rubra, Olibanum, Rhizoma Curcumae Longae are made Synsepalum duleificum leaf compound in the ratio of 36%, 28%, 18%, 18% respectively;
The acetone soln of Synsepalum duleificum leaf compound and 40% is mixed by weight 1:18, makes mixture, this mixture is heated to about 45 DEG C, supersound extraction 50 minutes, make Synsepalum duleificum leaf compound and extract solution;
By Synsepalum duleificum leaf compound and extraction solution coarse filtration, make compound recipe Synsepalum duleificum leaf extract coarse filtration liquid;
By compound recipe Synsepalum duleificum leaf extract coarse filtration liquid fine straining, make compound recipe Synsepalum duleificum leaf extract fine straining liquid;
Compound recipe Synsepalum duleificum leaf extract fine straining liquid is concentrated, makes compound recipe Synsepalum duleificum leaf extract concentrated solution;
The butanediol aqueous solution of compound recipe Synsepalum duleificum leaf extract concentrated solution 1:20 is redissolved, makes compound recipe Synsepalum duleificum leaf extract redissolution thing;
By the sterilizing of compound recipe Synsepalum duleificum leaf extract redissolution thing, make compound recipe Synsepalum duleificum leaf extract.
Embodiment 3
Synsepalum duleificum leaf, Fructus Atriplicis Sibiricae, Rhizoma Curcumae Longae, the Radix Astragali are made Synsepalum duleificum leaf compound in the ratio of 42%, 26%, 16%, 16% respectively;
Mixed by weight 1:15 by the acetone soln of Synsepalum duleificum leaf compound and 80%, make mixture, this mixture is heated to about 30 DEG C, heat of stirring refluxes 1 hour, makes Synsepalum duleificum leaf compound and extracts solution;
By Synsepalum duleificum leaf compound and extraction solution coarse filtration, make compound recipe Synsepalum duleificum leaf extract coarse filtration liquid;
By compound recipe Synsepalum duleificum leaf extract coarse filtration liquid fine straining, make compound recipe Synsepalum duleificum leaf extract fine straining liquid;
Compound recipe Synsepalum duleificum leaf extract fine straining liquid is concentrated, makes compound recipe Synsepalum duleificum leaf extract concentrated solution;
The butanediol aqueous solution of compound recipe Synsepalum duleificum leaf extract concentrated solution 1:20 is redissolved, makes compound recipe Synsepalum duleificum leaf extract redissolution thing;
By the sterilizing of compound recipe Synsepalum duleificum leaf extract redissolution thing, make compound recipe Synsepalum duleificum leaf extract.
Embodiment 4
Synsepalum duleificum leaf, Radix Paeoniae Rubra, Fructus Atriplicis Sibiricae, Olibanum are made Synsepalum duleificum leaf compound in the ratio of 45%, 25%, 15%, 15% respectively;
Mixed by weight 1:17 by the alcoholic solution of Synsepalum duleificum leaf compound and 50%, make mixture, this mixture is heated to about 45 DEG C, heat of stirring backflow supersound extraction 0.8 hour, makes Synsepalum duleificum leaf compound and extracts solution;
By Synsepalum duleificum leaf compound and extraction solution coarse filtration, make compound recipe Synsepalum duleificum leaf extract coarse filtration liquid;
By compound recipe Synsepalum duleificum leaf extract coarse filtration liquid fine straining, make compound recipe Synsepalum duleificum leaf extract fine straining liquid;
Compound recipe Synsepalum duleificum leaf extract fine straining liquid is concentrated, makes compound recipe Synsepalum duleificum leaf extract concentrated solution;
The butanediol aqueous solution of compound recipe Synsepalum duleificum leaf extract concentrated solution 1:20 is redissolved, makes compound recipe Synsepalum duleificum leaf extract redissolution thing;
By the sterilizing of compound recipe Synsepalum duleificum leaf extract redissolution thing, make compound recipe Synsepalum duleificum leaf extract.
Embodiment 5
Synsepalum duleificum leaf, Olibanum, Rhizoma Curcumae Longae, the Radix Astragali are made Synsepalum duleificum leaf compound in the ratio of 50%, 24%, 14%, 12% respectively;
Mixed by weight 1:12 by the alcoholic solution of Synsepalum duleificum leaf compound and 90%, make mixture, this mixture is heated to about 35 DEG C, heat of stirring refluxes 0.6 hour, makes Synsepalum duleificum leaf compound and extracts solution;
By Synsepalum duleificum leaf compound and extraction solution coarse filtration, make compound recipe Synsepalum duleificum leaf extract coarse filtration liquid;
By compound recipe Synsepalum duleificum leaf extract coarse filtration liquid fine straining, make compound recipe Synsepalum duleificum leaf extract fine straining liquid;
Compound recipe Synsepalum duleificum leaf extract fine straining liquid is concentrated, makes compound recipe Synsepalum duleificum leaf extract concentrated solution;
The butanediol aqueous solution of compound recipe Synsepalum duleificum leaf extract concentrated solution 1:20 is redissolved, makes compound recipe Synsepalum duleificum leaf extract redissolution thing;
By the sterilizing of compound recipe Synsepalum duleificum leaf extract redissolution thing, make compound recipe Synsepalum duleificum leaf extract.
Formula of the present invention is not limited to embodiment 1 ~ embodiment 5, and research finds, adopts other the combination in the scope of the invention also to have similar effect.
Embodiment 6: a kind of preparation of the whitening lotion containing compound recipe Synsepalum duleificum leaf extract
Above-mentioned compound recipe Synsepalum duleificum leaf extract is the arbitrary described compound recipe Synsepalum duleificum leaf extract of embodiment 1 ~ embodiment 5.
Embodiment 7: a kind of preparation of the cleawhite over-night essences containing compound recipe Synsepalum duleificum leaf extract
Above-mentioned compound recipe Synsepalum duleificum leaf extract is the arbitrary described compound recipe Synsepalum duleificum leaf extract of embodiment 1 ~ embodiment 5.
Embodiment 8: a kind of preparation of the whitening cream containing compound recipe Synsepalum duleificum leaf extract
Above-mentioned compound recipe Synsepalum duleificum leaf extract is the arbitrary described compound recipe Synsepalum duleificum leaf extract of embodiment 1 ~ embodiment 5.
Embodiment 9: a kind of preparation of the cleawhite pack containing compound recipe Synsepalum duleificum leaf extract
Above-mentioned compound recipe Synsepalum duleificum leaf extract is the arbitrary described compound recipe Synsepalum duleificum leaf extract of embodiment 1 ~ embodiment 5.
Embodiment 10: a kind of preparation of the whitening gel containing compound recipe Synsepalum duleificum leaf extract
Above-mentioned compound recipe Synsepalum duleificum leaf extract is the arbitrary described compound recipe Synsepalum duleificum leaf extract of embodiment 1 ~ embodiment 5.
Embodiment 11: a kind of preparation of the whitening emulsion containing compound recipe Synsepalum duleificum leaf extract
Above-mentioned compound recipe Synsepalum duleificum leaf extract is the arbitrary described compound recipe Synsepalum duleificum leaf extract of embodiment 1 ~ embodiment 4.
Carry out index test to the compound recipe Synsepalum duleificum leaf extract of embodiment 1-embodiment 5 gained 5%, testing result is all within the scope of following form.
| Testing index | Testing result |
| Flavones content/mgmL-1 | 0.39～0.43 |
| DPPH clearance rate/% | 81～93 |
| Tyrosinase inhibition rate/% | 84～93 |
| Hyaluronic acid suppression ratio/% | 57～68 |
Adopt the beneficial effect of above composite and process program: the present invention is by Synsepalum duleificum leaf and Radix Paeoniae Rubra, Fructus Atriplicis Sibiricae, the Radix Astragali, Rhizoma Curcumae Longae, any three kinds in Olibanum are carried out composite, have antioxidant activity and tyrosinase inhibitory activity concurrently, compensate for the deficiency that Synsepalum duleificum leaf only has stronger antioxidant activity, Synsepalum duleificum leaf compound is passed through pretreatment by the present invention, heat of stirring reflux, extract, coarse filtration, fine straining, concentrated, redissolve, compound recipe Synsepalum duleificum leaf extract is made in the process of sterilizing, its flavones content is higher, tyrosinase inhibition rate is also higher, can be used as white-skinned face function composition to be applied in cosmetics.
The active matter flavones content of compound recipe Synsepalum duleificum leaf extract product of the present invention detects
Compound recipe Synsepalum duleificum leaf extract crude drug mass concentration is 0.05g/mL, and flavones content testing result is as shown in table 1.
Table 1 compound recipe Synsepalum duleificum leaf extract flavones content
Remarks: flavones content detects and adopts NaNO
2-Al (NO
3)
3colorimetry.
One, the efficacy assessment of compound recipe Synsepalum duleificum leaf extract product of the present invention
1, tyrosinase inhibition rate
Participate in B16 cell and mainly contain three kinds of enzymes: tryrosinase, dopachrome interconvertible enzyme and dihydroxy indole carboxylic oxidase.Tryrosinase belongs to oxidoreductase, is the major rate-limiting enzyme of B16 cell, and tryrosinase and its substrate TYR catalytic reaction can occur.
After with the addition of the inhibiting reagent of tyrosinase activity in experimental system, can inhibitory action be produced to catalytic reaction, add before and after reagent in the light absorption value at 475nm place by measuring, evaluating the suppression ratio of reagent to tyrosinase activity.Under different tested concentration, the tyrosinase inhibition rate experimental result of compound recipe Synsepalum duleificum leaf extract is as shown in table 2.
Table 2 compound recipe Synsepalum duleificum leaf extract tyrosinase inhibition rate experimental result
As shown in Table 2, compound recipe Synsepalum duleificum leaf extract has good tyrosinase inhibition rate, and when tested concentration is 1.0%, tyrosinase inhibition rate reaches about 80%, shows that compound recipe Synsepalum duleificum leaf extract has good white-skinned face function at biochemistry level.And under identical concentration, the suppression ratio of compound recipe Synsepalum duleificum leaf extract tryrosinase is lower than commercially available competing product, and in tested concentration range, the tyrosinase inhibition rate of commercially available competing product all reaches about 100%.
2, hyaluronic acid enzyme inhibition rate
Hyaluronic acid enzyme inhibition rate size has reacted the antiinflammatory vigor of sample to a certain extent, and suppression ratio is higher, and anti-inflammatory activity is stronger.By building vitro inhibition hyaluronidase experimental system, assessment compound recipe Synsepalum duleificum leaf extract is to the suppression ratio of hyaluronidase.Under different tested concentration, compound recipe Synsepalum duleificum leaf extract suppresses hyaluronidase experimental result as shown in table 3.
Table 3 compound recipe Synsepalum duleificum leaf extract hyaluronic acid enzyme inhibition rate experimental result
As shown in Table 3, compound recipe Synsepalum duleificum leaf extract has certain hyaluronic acid enzyme inhibition rate, shows that it has certain anti-inflammatory activity.And under identical tested concentration, compound recipe Synsepalum duleificum leaf extract hyaluronidase inhibitor is all better than commercially available competing product.
3, skin microcirculation experiment is improved
Skin microcirculation blood flow is medically widely used.Within the specific limits, skin microcirculation blood flow shows that more greatly Skin Cell has good self renewal and regeneration capacity.The compound recipe Synsepalum duleificum leaf extract prepared is added in cream frost substrate, add concentration 5%, adopt " laser-Doppler blood flow tester " to analyze tested volunteer and use the situation of change improving skin microcirculation blood flow before and after the cream frost of compound recipe Synsepalum duleificum leaf extract.
Tested volunteer 15 people, in age 25-35 year, controls relative humidity (30 ± 5%) and temperature (25 ± 1 DEG C) in experimentation.Result shows, and compound recipe Synsepalum duleificum leaf extract has effect of good improvement skin microcirculation blood flow.Participate in experiment 15 volunteers, the skin microcirculation blood flow having 11 tested volunteers obtains effective raising, and effective percentage reaches 73%.In experiment, all 15 tested volunteer's skins all have no adverse reaction.
4, melanocyte test
Inquire into the compound recipe Synsepalum duleificum propagation of leaf extract sample on melanocyte and the impact of whitening function.Utilize B16 melanoma cell, working sample is on the impact of melanocyte proliferation function, and the impact on melanocyte synthesis of melanin.
(1) mensuration (mtt assay) of cell proliferation rate
Cultivate Murine melanoma B16 cells, treat that Growth of Cells arrives logarithmic (log) phase, use 0.25% trypsinization, adjustment cell concentration is 5 × 10
4individual/mL, is inoculated in 96 orifice plates, every hole 100 μ L.Be placed in after incubator cultivates 24h, abandon supernatant, add pastille culture medium, each sample three concentration groups.Each concentration establishes 3 multiple holes, and matched group does not add medicine, replaces corresponding solvent.After hatching 72h, every hole adds the MTT solution 20 μ L of 5g/L, abandons supernatant after 4h, and every hole adds cell pyrolysis liquid 100 μ L, and continue overnight incubation, microplate reader detects each hole absorbance, and wavelength is 570nm.
Cell proliferation inhibition rate %=(1-medicine group average absorbance value/matched group average absorbance value) × 100
(2) mensuration of B16 cell suppression ratio
By B16 melanocyte with 20 × 10
4the density of individual/mL is inoculated in 24 plates.Every hole 1mL, after hatching 24h, abandons supernatant, adds medicine to be measured (sample effect final concentration is with aforementioned), after drug effect 72h, abandons supernatant, and PBS washs, and every hole adds the 1mol/LNaOH solution 500 μ L containing 10%DMSO, concussion 5min.Each hole light absorption value under measurement 490nm.
B16 cell suppression ratio %=[1-(medicine hole 490nm absorbance/medicine hole 570nm absorbance)/(control wells 490nm absorbance/control wells 570nm absorbance)] × 100
After adding variable concentrations compound recipe Synsepalum duleificum leaf extract tested material, sample to melanocyte melanin synthesis suppression ratio and cell proliferation inhibition rate experimental result as shown in table 4.
Table 4 variable concentrations compound recipe Synsepalum duleificum leaf extract sample is to the inhibited proliferation of B16 cell and melanin nucleus formation
Conclusion: after adding the tested material of variable concentrations, compound recipe Synsepalum duleificum leaf extract sample has the effect of good check melanin cell synthesis of melanin, in tested concentration range (0.5%-2.0%), B16 cell suppression ratio is at about 30%-40%, show that compound recipe Synsepalum duleificum leaf extract has good white-skinned face function at cellular level, and the impact of on cell proliferation is lower than commercially available competing product.
Two, the safety evaluation of compound recipe Synsepalum duleificum leaf extract product of the present invention
1, erythrocyte hemolysis test
Erythrocyte hemolysis rate to a certain extent can the zest of response sample, and hemolysis rate is higher, and potential zest is larger.Compound recipe Synsepalum duleificum leaf extract extracting solution erythrocyte hemolysis rate experimental result is as shown in table 5 below.
Table 5 compound recipe Synsepalum duleificum leaf extract erythrocyte hemolysis rate experimental result
As shown in Table 5, compound recipe Synsepalum duleificum leaf extract extracting solution has extremely low erythrocyte hemolysis rate in the concentration range of 10%, shows that it has good safety.
2, mice repeatedly Skin Irritation Test
By mice repeatedly Skin Irritation Test, to determine and whether evaluation compound recipe Synsepalum duleificum leaf extract has stimulation or corrosiveness and degree thereof to mammal skin local.
Skin irritation/corrosion test method that method specifies with reference to 2007 editions cosmetics health specifications.By tested material once (or repeatedly) be coated on the skin of animal subject, in predetermined time interval, observe the degree of animal skin local irritant effect and mark.Adopt own control, to evaluate the stimulation of tested material to skin.The acute skin irritation experimental observation time limit should be enough to reversibility or the irreversibility of evaluating this effect.
Result of the test shows, and smear 14 days given the test agent (adding compound recipe Synsepalum duleificum leaf extract extracting solution 5% and 10% respectively) continuously, erythema and edema phenomenon all do not appear in mouse skin.
Result illustrates, the safety of compound recipe Synsepalum duleificum leaf extract is good, nonirritant.
3, human closed formula patch test
Human body patch test method is with reference to " cosmetic hygienic practice (2007 editions) ", and preparation carries out closed patch containing the cream frost of the compound recipe Synsepalum duleificum leaf extract extracting solution of 1.0% and 5.0%, investigates sample causes untoward reaction probability to human body.
Skin adverse reaction classification is as shown in table 7 below.
The classification of table 6 skin adverse reaction
According to table 6, observation experimenter removes dermoreaction situation when speckle examination device 30min, 24h and 48h.
Result of the test is as shown in following table 7-table 9.
Speckle examination device 30min observed result removed by table 7
Speckle examination device 24h observed result removed by table 8
Speckle examination device 48h observed result removed by table 9
According to the requirement of 2007 " cosmetics health specifications ", judgement sample 1, sample 2 pairs of human body skins have no adverse reaction.(wherein, sample 1 is the cream frost of the compound recipe Synsepalum duleificum leaf extract extracting solution of interpolation 1.0%, and sample 2 is the cream frost of the compound recipe Synsepalum duleificum leaf extract extracting solution of interpolation 5.0%.)
Conclusion: the cream frost adding the compound recipe Synsepalum duleificum leaf extract of 1.0%, 5.0% two concentration, have no adverse reaction to human body skin, safety is good.
4, Phototoxicity experiment
Adopt external 3T3 red untake Phototoxicity experiment to assess compound recipe Synsepalum duleificum leaf extract and whether there is potential phototoxicity.
Whether there is phototoxicity by calculating photostimulation factor PIF and average light effect MPE comprehensive assessment sample, judgment criteria is as follows: as PIF<2.0 or MPE<0.1: expection is without phototoxicity; As PIF>2.0 and <5.0 or MPE>0.1 and <0.15: expection may exist phototoxicity; As PIF>5.0 or MPE>0.15: expection exists phototoxicity.
Test adopts 96-well culture plate to carry out.Balb/c3T3 cell good for growth conditions is inoculated in 96 porocyte culture plates (by test product and each 2 blocks of plates of positive reference substance), makes the cell density in every hole be about 1.0 × 10
4individual/hole.Cultivate after 24 hours, add the HBSS solution being subject to test product drug-delivery preparation and positive control hydrochloric acid chlorine piperazine (CPZ) containing variable concentrations, arrange blank and excipient/Vehicle controls simultaneously, the multiple hole of each process 6, often kind by test product 2 blocks of plates.After humidification cultivates 1 hour, carry out illumination and non-photo-irradiation treatment respectively, then change fresh culture fluid and continue to cultivate 18-22 hour.Cultivate and terminate precontract 3 hours, the DMEM culture fluid added containing dimethyl diaminophenazine chloride serum-free continues to cultivate.After cultivation terminates, every hole adds dimethyl diaminophenazine chloride eluent, is placed in agitator quick oscillation until form uniform solution.Under 540nm, the OD value (OD540) in each hole is measured with spectrophotometer.
With the Phototox computed in software photostimulation factor (PIF) value.In test, the OD of excipient or solvent control group
540numerical value is all greater than 0.4 and positive reference substance hydrochloric acid chlorobenzene piperazine IC
50(+UV) within the scope of 0.1-3.0 μ g/ml, IC
50(-UV) is within the scope of 7.0 – 90.0 μ g/ml, and PIF ﹥ 6, show that pilot system is set up.
In test, during application of sample and after process terminates, each concentration group of sample all do not observe by test product Precipitation and morphocytology abnormal; With Phototox computed in software result of the test, the PIF value of sample is * 1, that is: tested material does not also show cytotoxicity reaching the maximum concentration value of permission (1000.0 μ g/mL); Thus tested material sample is without potential phototoxicity; The MPE value of sample is also all less than 0.1, further illustrates the expection of tested material sample without phototoxicity.
Table 10 compound recipe Synsepalum duleificum leaf extract Phototoxicity experiment result
Remarks: * 1: also do not show cytotoxicity in the maximum concentration value (1000.0 μ g/mL) reaching permission; +: there is phototoxicity in expection;-: expection is without phototoxicity.
Therefore, under this experimental condition, the external 3T3 red untake Phototoxicity experiment of given the test agent is negative reaction, and expection is without phototoxicity.
5, Skin allergy test
Skin allergy test is with reference to cosmetics health specification (2007 editions), and adopt partial closure to be coated with skin test and test (BuehlerTest, BT) method, whether that determines compound recipe Synsepalum duleificum leaf extract causes skin allergic reaction and degree.The scoring of allergy test dermoreaction is as shown in table 11.
The scoring of table 11 allergy test dermoreaction
Evaluation of result: when dermoreaction integration >=2 appear in tested material treated animal, is judged to this animal and occurs that skin allergic reaction is positive, judge the sensitization intensity of tested material by table 12.
Table 12 sensitization rate is classified
Compound recipe Synsepalum duleificum leaf extract (under 5%, 10% two concentration) is as shown in table 13 to guinea pig skin allergic provocative test result.Result shows, and guinea pig skin has no allergy, and sensitization rate is 0, animal activity and Normal-weight.
Table 13 compound recipe Synsepalum duleificum leaf extract is to the allergic provocative test result of guinea pig skin
Remarks:
Vegetable oil is negative control;
2,4 dinitro-chloro-benzenes are positive control;
Sample 1 is 5% compound recipe Synsepalum duleificum leaf extract solution;
Sample 2 is 10% compound recipe Synsepalum duleificum leaf extract solution.
Three, conclusion
1, efficacy assessment conclusion
Through the both effectiveness assessment experiment of system, effect of compound recipe Synsepalum duleificum leaf extract extracting solution is summarized as follows:
(1) compound recipe Synsepalum duleificum leaf extract extracting solution has good restraint of tyrosinase activity, and tested concentration is 1.0% time, and namely tyrosinase inhibition rate reaches about 80%.
(2) compound recipe Synsepalum duleificum leaf extract extracting solution has certain anti-inflammatory activity, and tested concentration is 5.0% time, and hyaluronic acid enzyme inhibition rate is about 65%, and the anti-inflammatory activity of compound recipe Synsepalum duleificum leaf extract is better than commercially available competing product.
(3) improve the display of microcirculation experimental result, compound recipe Synsepalum duleificum leaf extract has effect of good improvement skin microcirculatory blood flow.
(4) melanocyte result of the test display, compound recipe Synsepalum duleificum leaf extract has certain B16 cell suppression ratio under tested concentration 0.5%, 1% and 2% 3 tested concentration, can ensure that the relative motility rate of melanocyte is more than 80% simultaneously.Show, compound recipe Synsepalum duleificum leaf extract has certain white-skinned face function at cellular level.
2, safety evaluation conclusion
By erythrocyte hemolysis experiment, mice repeatedly Skin Irritation Test, human body patch test, Phototoxicity experiment and Skin allergy test, the safety of assessment compound recipe Synsepalum duleificum leaf extract extracting solution, five safety experiment results unanimously show, compound recipe Synsepalum duleificum leaf extract extracting solution is safe, non-stimulated, to have no adverse reaction generation to human body.
Above-described embodiment is only be described the preferred embodiment of the present invention; not scope of the present invention is limited; under not departing from the present invention and designing the prerequisite of spirit; the various distortion that those of ordinary skill in the art make technical scheme of the present invention and improvement, all should fall in protection domain that claims of the present invention determines.
Claims (7)
1. a compound recipe Synsepalum duleificum leaf extract, it is characterized in that: described compound recipe Synsepalum duleificum leaf extract is prepared from herbal raw material, described herbal raw material is made up of major ingredient and adjuvant, described major ingredient is Synsepalum duleificum leaf, percentage by weight is 30% ~ 50%, described adjuvant is the combination of any three kinds in Radix Paeoniae Rubra, Fructus Atriplicis Sibiricae, Olibanum, Rhizoma Curcumae Longae, the Radix Astragali, and percentage by weight is respectively 20% ~ 30%, 10% ~ 30% and 10% ~ 20%.
2. compound recipe Synsepalum duleificum leaf extract according to claim 1, is characterized in that: the preparation method of described compound recipe Synsepalum duleificum leaf extract comprises the steps:
(1) described herbal raw material is prepared in proportion;
(2) pretreatment: carry out the broken process of ovendry power after being chosen by the foreign body in described herbal raw material, obtain herbal raw material powder, by four kinds of herbal powder mix homogeneously, makes Synsepalum duleificum leaf compound;
(3) extract: described Synsepalum duleificum leaf compound and solvent are mixed by weight 1:11 ~ 18, extracts 0.5 ~ 1 hour at the temperature of 26 ~ 45 DEG C, obtain extraction solution; Described solvent is pure water, the alcoholic solution of 40% ~ 95% or the acetone soln of 40% ~ 95%;
(4) coarse filtration: by described extraction solution coarse filtration, obtains compound recipe Synsepalum duleificum leaf extract coarse filtration liquid;
(5) fine straining: by described compound recipe Synsepalum duleificum leaf extract coarse filtration liquid fine straining, obtain compound recipe Synsepalum duleificum leaf extract fine straining liquid;
(6) concentrated: by described compound recipe Synsepalum duleificum leaf extract fine straining liquid, to obtain compound recipe Synsepalum duleificum leaf extract concentrated solution;
(7) redissolve: adopted by described compound recipe Synsepalum duleificum leaf extract concentrated solution the butanediol aqueous solution of 1:20 to redissolve, obtain compound recipe Synsepalum duleificum leaf extract redissolution thing;
(8) sterilizing, by the sterilizing of described compound recipe Synsepalum duleificum leaf extract redissolution thing, obtains compound recipe Synsepalum duleificum leaf extract.
3. compound recipe Synsepalum duleificum leaf extract according to claim 2, is characterized in that: be extracted as stirring and refluxing described in step (3) and extract or supersound extraction.
4. the application of compound recipe Synsepalum duleificum leaf extract in skin-lightening cosmetic in claims 1 to 3 described in any one.
5. application according to claim 4, is characterized in that: in described skin-lightening cosmetic, and described compound recipe Synsepalum duleificum leaf extract accounts for 0.2% ~ 8%.
6. application according to claim 5, is characterized in that: also comprise one or more in solvent, emollient, solubilizing agent, antioxidant, pH adjusting agent, penetrating agent, emulsion stabilizer, antiseptic, wetting agent, thickening agent, nertralizer, chelating agen, surfactant, emulsifying agent, essence, coloring agent, sunscreen, skin conditioning agent, active matter and functional additive in described skin-lightening cosmetic.
7. application according to claim 6, is characterized in that: described skin-lightening cosmetic is astringent, facial film, facial cream, gel, emulsion, essence.
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| CN110772456A (en) * | 2019-11-04 | 2020-02-11 | 中国热带农业科学院南亚热带作物研究所 | Miracle fruit leaf compound and application thereof in anti-melanin formation product |
| CN116019755A (en) * | 2023-02-14 | 2023-04-28 | 太和康美(北京)中医研究院有限公司 | Method for reducing or removing odor of plant extract and application thereof |
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| CN107375180A (en) * | 2017-08-11 | 2017-11-24 | 北京斯利安药业有限公司 | A kind of composite skin care product, facial mask and preparation method thereof |
| CN107440932A (en) * | 2017-08-11 | 2017-12-08 | 北京斯利安药业有限公司 | A kind of composite skin care product containing folic acid, facial mask and preparation method thereof |
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| CN108553390A (en) * | 2018-07-01 | 2018-09-21 | 佛山市芊茹化妆品有限公司 | A kind of preparation method and applications of compound skin-whitening composition |
| CN110772456A (en) * | 2019-11-04 | 2020-02-11 | 中国热带农业科学院南亚热带作物研究所 | Miracle fruit leaf compound and application thereof in anti-melanin formation product |
| CN110772456B (en) * | 2019-11-04 | 2022-08-23 | 中国热带农业科学院南亚热带作物研究所 | Miracle fruit leaf compound and application thereof in anti-melanin formation product |
| CN116019755A (en) * | 2023-02-14 | 2023-04-28 | 太和康美(北京)中医研究院有限公司 | Method for reducing or removing odor of plant extract and application thereof |
| CN116019755B (en) * | 2023-02-14 | 2023-10-10 | 太和康美(北京)中医研究院有限公司 | Method for reducing or removing odor of plant extract and application thereof |
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