CN105250167A - Compound synsepalum dulcificum leaf extract and application thereof in anti-aging cosmetics - Google Patents
Compound synsepalum dulcificum leaf extract and application thereof in anti-aging cosmetics Download PDFInfo
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- 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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Abstract
The invention discloses a compound synsepalum dulcificum leaf extract. The compound synsepalum dulcificum leaf extract is prepared from Chinese herbal medicine raw materials, the main material is synsepalum dulcificum leaf, and the auxiliary material is a combination of any three of radix paeoniae rubra, tribulus terrestris, astragalus, rhodiola rosea and scutellaria baicalensis. The invention also discloses a preparation method of the compound synsepalum dulcificum leaf extract. The method includes: preparation of Chinese herbal medicine raw materials, pretreatment, extraction, rough filtration, fine filtration, concentration, re-dissolution and sterilization. The invention also discloses application of the compound synsepalum dulcificum leaf extract in cosmetics with anti-aging efficacy. The compound synsepalum dulcificum leaf extract provided by the invention has antioxidant activity and tyrosinase inhibitory activity, makes up the drawback that synsepalum dulcificum leaf only has strong antioxidant activity, and can be applied as an anti-aging functional component in cosmetics.
Description
Technical field
The present invention relates to cosmetic field, particularly relate to a kind of preparation method of compound recipe Synsepalum duleificum leaf extract and the application in anti-aging cosmetics 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 is to provide and a kind ofly has the compound recipe Synsepalum duleificum leaf extract of antioxidant activity and tyrosinase inhibitory activity and the application in anti-aging cosmetics thereof concurrently, and provides the extracting method of this compound recipe Synsepalum duleificum leaf extract.
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%, described adjuvant is the combination of any three kinds in Radix Paeoniae Rubra, Fructus Atriplicis Sibiricae, the Radix Astragali, Radix Rhodiolae, Radix Scutellariae, 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 1:19 ~ 26, extracts 1.5 ~ 2 hours at the temperature of 46 ~ 59 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 the cosmetics with senile-resistant efficacy.
Application of the present invention, wherein, in described cosmetics, 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 cosmetics.
Application of the present invention, wherein, described cosmetics are astringent, facial film, facial cream, gel, emulsion, essence.
The preparation method difference from prior art of compound recipe Synsepalum duleificum leaf extract of the present invention is:
Synsepalum duleificum leaf has good antioxidant activity, but there is no the function of restraint of tyrosinase activity, want to play its outstanding anti-ageing effect of waiting for a long time, 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 senile-resistant efficacy.
The present invention is by Synsepalum duleificum leaf and Radix Paeoniae Rubra, Fructus Atriplicis Sibiricae, the Radix Astragali, Radix Rhodiolae, Radix Scutellariaes etc. carry 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, DPPH free radical scavenging activity, ultra-oxygen anion free radical clearance rate, tyrosinase inhibition rate is also higher, can be used as senile-resistant efficacy composition to be 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 1:19, makes mixture, this mixture is heated to about 46 DEG C, supersound extraction 1.5 hours, 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, Radix Rhodiolae, Radix Scutellariae 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 1:26, makes mixture, this mixture is heated to about 59 DEG C, supersound extraction 2 hours, 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, Radix Rhodiolae, Radix Scutellariae, the Radix Astragali are made Synsepalum duleificum leaf compound in the ratio of 42%, 26%, 16%, 16% respectively;
Mixed by weight 1:20 by the acetone soln of Synsepalum duleificum leaf compound and 80%, make mixture, this mixture is heated to about 50 DEG C, heat of stirring refluxes 1.6 hours, 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, Fructus Atriplicis Sibiricae, Radix Scutellariae, the Radix Astragali are made Synsepalum duleificum leaf compound in the ratio of 45%, 25%, 15%, 15% respectively;
Mixed by weight 1:25 by the alcoholic solution of Synsepalum duleificum leaf compound and 50%, make mixture, this mixture is heated to about 55 DEG C, heat of stirring backflow supersound extraction 1.8 hours, 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, Radix Paeoniae Rubra, Radix Rhodiolae, the Radix Astragali are made Synsepalum duleificum leaf compound in the ratio of 50%, 24%, 14%, 12% respectively;
Mixed by weight 1:24 by the alcoholic solution of Synsepalum duleificum leaf compound and 90%, make mixture, this mixture is heated to about 52 DEG C, heat of stirring refluxes 1.7 hours, 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 with the astringent of senile-resistant efficacy 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 with the essence of senile-resistant efficacy 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 with the facial cream of senile-resistant efficacy 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 with the facial film of senile-resistant efficacy 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 with the gel of senile-resistant efficacy 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 with the emulsion of senile-resistant efficacy 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, Radix Rhodiolae, any three kinds in Radix Scutellariae 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, DPPH free radical scavenging activity, ultra-oxygen anion free radical clearance rate, tyrosinase inhibition rate is also higher, can be used as senile-resistant efficacy 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, DPPH free radical scavenging activity
DPPH is a kind of stable organic free radical, be 517nm at visible region maximum absorption band, in alcoholic solution, each DPPH molecule can generate in the solution one stable containing nitrogen free radical, there is typical purple, when it is with when providing the free radical scavenger effect of 1 electronics, generate colourless product, the typical purple of solution is shoaled, and its fading extent becomes stoichiometric relationship with sharing electron number.Therefore, available spectrophotography carries out quantitative analysis, detects free radical scavenging situation, thus the ability of the scavenging free radicals of assess sample.Research shows, DPPH clearance rate is higher, and antioxidant activity is stronger.Under different tested concentration, the DPPH clearance rate experimental result of compound recipe Synsepalum duleificum leaf extract is as shown in table 2.
Table 2 compound recipe Synsepalum duleificum leaf extract DPPH free radical scavenging experimental result
As shown in Table 2, compound recipe Synsepalum duleificum leaf extract has good DPPH free radical scavenging effect, tested concentration is 2.0% time, namely DPPH free radical scavenging activity reaches about 88%, show that compound recipe Synsepalum duleificum leaf extract has good antioxidant activity at biochemistry level, and the antioxidant activity of compound recipe Synsepalum duleificum leaf extract is better than commercially available competing product.
2, 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 3.
Table 3 compound recipe Synsepalum duleificum leaf extract tyrosinase inhibition rate experimental result
As shown in Table 3, 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 senile-resistant efficacy 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%.
3, 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 4.
Table 4 compound recipe Synsepalum duleificum leaf extract hyaluronic acid enzyme inhibition rate experimental result
As shown in Table 4, 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.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 6 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 antioxidant activity, and tested concentration is 2.0% time, and namely DPPH clearance rate reaches about 88%, and the antioxidant activity of Synsepalum duleificum leaf is better than commercially available competing product.
(2) 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%.
(3) 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.
Show, compound recipe Synsepalum duleificum leaf extract has certain senile-resistant efficacy 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, the Radix Astragali, Radix Rhodiolae, Radix Scutellariae, 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 1:19 ~ 26, extracts 1.5 ~ 2 hours at the temperature of 46 ~ 59 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 the cosmetics with senile-resistant efficacy in claims 1 to 3 described in any one.
5. application according to claim 4, is characterized in that: in described cosmetics, 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 cosmetics.
7. application according to claim 6, is characterized in that: described cosmetics are astringent, facial film, facial cream, gel, emulsion, essence.
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| CN201510632159.XA CN105250167A (en) | 2015-09-29 | 2015-09-29 | Compound synsepalum dulcificum leaf extract and application thereof in anti-aging cosmetics |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
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Application publication date: 20160120 |