CN108553390A - A kind of preparation method and applications of compound skin-whitening composition - Google Patents
A kind of preparation method and applications of compound skin-whitening composition Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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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- A61K8/9789—Magnoliopsida [dicotyledons]
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- A61K8/00—Cosmetics or similar toiletry preparations
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- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
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- 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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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
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Abstract
The invention discloses a kind of compound skin-whitening composition mainly having both anti-inflammatory, antioxidant activity and tyrosinase inhibitory activity made from Caesalpinia spinosaKuntze and its applications in cosmetics, and the extracting method of the compound skin-whitening composition is provided, extracting method includes herbal raw material pretreatment, extraction, filtering, concentration, redissolution and sterilizing.The invention also discloses application of the compound skin-whitening composition in cosmetics.Compound skin-whitening composition of the present invention has both antioxidant activity and tyrosinase inhibitory activity, can be used as whitening anti-aging functional component applied in cosmetics.
Description
Technical field
The present invention relates to cosmetic fields, are specifically related to a kind of preparation method and applications of compound skin-whitening composition.
Background technology
Caesalpinia spinosaKuntze system Caesalpiniaceae Caesalpinia, Classification system are Caesalpinia spinosa Kuntze, are short weeks
Phase preciousness economic tree is grown on South America northwestward, and locality claims tower to draw (TARA), its beans shell 47-55% of tannin containing mysorethorn,
It is the important source material of a variety of chemical industry of production, medical product, there is extensive industrial use；Its seed draws polysaccharide 22- containing tower
34%, it can be used for producing tara gum (Taragum), tara gum has in food, medicine, weaving, papermaking and oil drilling etc.
Important use；Its embryo 40-60% containing crude protein, crude fat 14% or so can produce low fat legumin, the fat contained by embryo
In fat acid, linoleic acid accounts for 45% or so, and linoleic acid is a kind of important medical material.Though Caesalpinia spinosaKuntze is applied in cosmetic field
Have not been reported, but because its embryo is rich in polysaccharide, crude protein, trace element is abundant, wherein phosphorus, iron, zinc content compare soybean
Height, and it is nontoxic, there is great potential application value in cosmetics.
Invention content
The technical problem to be solved in the present invention is to provide one kind mainly having both anti-inflammatory, antioxidant activity made from Caesalpinia spinosaKuntze
Compound skin-whitening composition with tyrosinase inhibitory activity and its application in cosmetics, and the compound skin-whitening composition is provided
Extracting method.
A kind of preparation method of compound skin-whitening composition, includes the following steps：
(1) prepare herbal raw material in proportion：The herbal raw material is made of major ingredient and auxiliary material, and the major ingredient is thorn cloud
Real fruit, weight percent 30-50%, auxiliary material are short cut tobacco orchid root, the hard English walnut seeds of Bai Er, climbing entada stem rattan, weight percent
Respectively 20-30%, 10-30% and 10-20%；
(2) it pre-processes：Climbing entada stem rattan is first taken, impurity is removed, sets in frying container, heated with slow boiling, fries to color and adds
It is deep to yellow black, when aromatic effusion, taking-up is dried in the air cool, and the used time crushes；It then will be different in other above-mentioned three tastes herbal raw materials
Object carries out dries pulverizing processing after choosing, and obtains herbal raw material powder；Four kinds of herbal powders are uniformly mixed again, thorn cloud is made
Real fruit compound；
(3) it extracts：The Caesalpinia spinosaKuntze fruit compound and solvent are mixed and extracted, extraction solution is obtained；
(4) it filters：The extraction solution is filtered, extract filtrate is obtained；
(5) it concentrates：The extract filtrate is concentrated, extract concentrated solution is obtained；
(6) it redissolves：The extract concentrated solution is redissolved, extract is obtained and redissolves object；
(7) it sterilizes, the extract is redissolved into object sterilizing, obtains compound skin-whitening composition.
The preparation method of compound skin-whitening composition of the present invention, wherein Caesalpinia spinosaKuntze fruit compound described in step (3)
With solvent by weight 1:(10-40) is mixed, and is extracted 1-3 hours at a temperature of 80-90 DEG C, is obtained extraction solution.
The preparation method of compound skin-whitening composition of the present invention, wherein filtering is 80-100 mesh described in step (4)
Filtering.
The preparation method of compound skin-whitening composition of the present invention, wherein redissolved described in step (6) and use 1:20
Dipropylene glycol aqueous solution redissolves.
The preparation method of compound skin-whitening composition of the present invention, wherein solvent described in step (3) is 60-90%
Ethanol solution or 60-90% acetone soln.
The preparation method of compound skin-whitening composition of the present invention, wherein step is extracted as being stirred at reflux described in (3)
Extraction or ultrasonic extraction.
Application of the compound skin-whitening composition of the present invention in cosmetics.
The dosage of a kind of cosmetics, including the compound skin-whitening composition, the compound skin-whitening composition is final production
The 1-10% of product weight.
Cosmetics of the present invention, wherein the dosage of the compound skin-whitening composition is the 2- of final products weight
8%.
Cosmetics of the present invention, wherein further include solvent in the cosmetics, emollient, solubilizer, anti-oxidant
Agent, pH adjusting agent, penetrating agent, emulsion stabilizer, preservative, moisturizer, thickener, neutralizer, chelating agent, surfactant,
One or more of emulsifier, essence, colorant, sun-screening agent, skin conditioning agent, active matter and functional additive.
Cosmetics of the present invention, wherein the cosmetics are toner, shower cream, face cleaning cream, facial mask, face cream, Gel
Curry, lotion, Essence, makeup remover or transparent soap.
The preparation method difference from prior art of compound skin-whitening composition of the present invention is：
Caesalpinia spinosaKuntze fruit has good anti-inflammatory, antioxidant activity, but the function of not inhibiting tyrosinase activity, to hair
Its outstanding anti-aging, Bearberry Extract are waved, need to consider to carry out compound extraction, it is anti-inflammatory, anti-oxidant to give full play to Caesalpinia spinosaKuntze fruit
While active, more excellent skin-care effect is carried.
About the selection of compound, according to theory of traditional Chinese medical science " dialectical, entirety, synthesis " and the monarch and his subjects' principles of formulating prescriptions " monarch, minister, assistant,
Make ", it is compounded, wherein determining that Caesalpinia spinosaKuntze fruit is the main status of " monarch ", the selection Chinese herbal medicine that disposition matches therewith is answered
With prescription, polysaccharide is obtained to maximize, crude protein, trace element is basis for estimation, and centre extracts the optimization design of technique,
It can be that skin supplements the nutrients to be desirably to obtain, and all kinds of cosmetics are applied to the best extracting solution of white-skinned face function, senile-resistant efficacy
In dosage form.
Caesalpinia spinosaKuntze fruit and short cut tobacco orchid root, the hard English walnut seeds of Bai Er, climbing entada stem rattan are compounded, are had both anti-by the present invention
Scorching, antioxidant activity and tyrosinase inhibitory activity, compensating for Caesalpinia spinosaKuntze fruit only has stronger anti-inflammatory, antioxidant activity not
Foot, the present invention pre-process climbing entada stem rattan by processing, and can remove toxic component therein, to prevent causing to stimulate to skin
Injury, must improve the yield of its active ingredient, by Caesalpinia spinosaKuntze fruit compound by crushing, stirring circumfluence distillation, through 80-
100 mesh filter, and can remove impurity and macromolecular, obtain active preferably useful small molecule ingredient, then concentrate, redissolve,
Compound skin-whitening composition is made in the processing of sterilizing, and flavones content is higher, DPPH free radical scavenging activities, ultra-oxygen anion free radical
Clearance rate, tyrosinase inhibition rate are also relatively high, can be used as whitening anti-aging functional component applied in cosmetics.
Specific implementation mode
To make the object, technical solutions and advantages of the present invention clearer, embodiment of the present invention will be made into one below
Step ground detailed description, embodiments of the present invention are not limited thereto, for not specifically specified technological parameter or condition, can refer to
Routine techniques carries out.
Short cut tobacco orchid of the present invention is the monocotyledon of Liliaceae Yucca, lives in the North America west and south, Latin
Scientific name is Yuccabrevifolia, is used as medicine with root.Play the role of anti-inflammatory；There is certain elimination to act on free radical simultaneously, has anti-
The effect of oxidation；There is facilitation to the decomposition of fat, can be used as diet products, in addition may be used as antiseptic, the suppression of mite allergy
Preparation.
The hard English walnuts of Bai Er of the present invention belong to Anacardiaceae plant, originate in South Africa, Classification system is
Sclerocaryabirrea (A.Rich) Hochst.subsp.caffra (Sond.) Kokwaro, seed are used as medicine.Fruit can be fresh
Food, is cooked jam, fruit juice and jelly, can also be used to alcoholic beverage of making wine and manufacture.The hard English walnut seed oil contents of Bai Er are 44-
The hard nut oil of 72%, Bai Er has unique nutty flavor, contains abundant unsaturated fatty acid, vitamin E and sterol and Huang
The extract of letones, bark and leaf contains antidiarrheal, anti-inflammatory, anti-corrosion, sterilization, anticoagulation, blood pressure lowering, hypoglycemic, anti-convulsion
Contraction and anti-oxidant equal substances.
Climbing entada of the present invention is Dou Ke Ke vine platymisciums, and Classification system is Entada phaseoloides (L.)
Merr., it is used as medicine with rattan and benevolence.It is slight bitter, puckery, it puts down, there is dispelling wind and eliminating dampness, effect promoting blood circulation and removing obstruction in channels.
Embodiment 1
Climbing entada stem rattan is first taken, impurity is removed, sets in frying container, heated with slow boiling, stir-fry to color burn to yellow black,
When aromatic effusion, cool, used time crushing of drying in the air is taken out；Then it is carried out after the foreign matter in other above-mentioned three tastes herbal raw materials being chosen
Dries pulverizing processing, obtains herbal raw material powder；
By Caesalpinia spinosaKuntze fruit, short cut tobacco orchid root, the hard English walnut seeds of Bai Er, climbing entada stem rattan respectively according to 30%, 30%, 20%,
Caesalpinia spinosaKuntze fruit compound is made in 20% ratio；
By Caesalpinia spinosaKuntze fruit compound and 60% ethanol solution by weight 1:10 are mixed, and mixture is made, by this
Mixture is heated to 90 DEG C, and the extraction solution of Caesalpinia spinosaKuntze fruit compound is made in ultrasonic extraction 1 hour；
80 mesh of extraction solution of Caesalpinia spinosaKuntze fruit compound is filtered, extract filtrate is made；
Extract filtrate is concentrated, extract concentrated solution is made；
By extract concentrated solution with 1:20 dipropylene glycol aqueous solution redissolves, and extract is made and redissolves object；
Extract is redissolved into object sterilizing, compound skin-whitening composition is made.
Embodiment 2
Climbing entada stem rattan is first taken, impurity is removed, sets in frying container, heated with slow boiling, stir-fry to color burn to yellow black,
When aromatic effusion, cool, used time crushing of drying in the air is taken out；Then it is carried out after the foreign matter in other above-mentioned three tastes herbal raw materials being chosen
Dries pulverizing processing, obtains herbal raw material powder；
By Caesalpinia spinosaKuntze fruit, short cut tobacco orchid root, the hard English walnut seeds of Bai Er, climbing entada stem rattan respectively press 40%, 20%, 30%,
Caesalpinia spinosaKuntze fruit compound is made in 10% ratio；
By Caesalpinia spinosaKuntze fruit compound and 90% acetone soln by weight 1:25 are mixed, and mixture is made, by this
Mixture is heated to 80 DEG C, and the extraction solution of Caesalpinia spinosaKuntze fruit compound is made in ultrasonic extraction 3 hours；
100 mesh of extraction solution of Caesalpinia spinosaKuntze fruit compound is filtered, extract filtrate is made；
Extract filtrate is concentrated, extract concentrated solution is made；
By extract concentrated solution with 1:20 dipropylene glycol aqueous solution redissolves, and extract is made and redissolves object；
Extract is redissolved into object sterilizing, compound skin-whitening composition is made.
Embodiment 3
Climbing entada stem rattan is first taken, impurity is removed, sets in frying container, heated with slow boiling, stir-fry to color burn to yellow black,
When aromatic effusion, cool, used time crushing of drying in the air is taken out；Then it is carried out after the foreign matter in other above-mentioned three tastes herbal raw materials being chosen
Dries pulverizing processing, obtains herbal raw material powder；
By Caesalpinia spinosaKuntze fruit, short cut tobacco orchid root, the hard English walnut seeds of Bai Er, climbing entada stem rattan respectively press 50%, 25%, 10%,
Caesalpinia spinosaKuntze fruit compound is made in 15% ratio；
By Caesalpinia spinosaKuntze fruit compound and 75% acetone soln by weight 1:25 are mixed, and mixture is made, by this
Mixture is heated to 85 DEG C, stirs heat reflux 2 hours, the extraction solution of Caesalpinia spinosaKuntze fruit compound is made；
90 mesh of extraction solution of Caesalpinia spinosaKuntze fruit compound is filtered, extract filtrate is made；
Extract filtrate is concentrated, extract concentrated solution is made；
By extract concentrated solution with 1:20 dipropylene glycol aqueous solution redissolves, and extract is made and redissolves liquid；
Extract is answered into solution sterilization, compound skin-whitening composition is made.
Embodiment 4
Climbing entada stem rattan is first taken, impurity is removed, sets in frying container, heated with slow boiling, stir-fry to color burn to yellow black,
When aromatic effusion, cool, used time crushing of drying in the air is taken out；Then it is carried out after the foreign matter in other above-mentioned three tastes herbal raw materials being chosen
Dries pulverizing processing, obtains herbal raw material powder；
By Caesalpinia spinosaKuntze fruit, short cut tobacco orchid root, the hard English walnut seeds of Bai Er, climbing entada stem rattan respectively press 45%, 25%, 15%,
Caesalpinia spinosaKuntze fruit compound is made in 15% ratio；
By Caesalpinia spinosaKuntze fruit compound and 70% ethanol solution by weight 1:30 are mixed, and mixture is made, by this
Mixture is heated to 88 DEG C, stirs heat reflux ultrasonic extraction 2.5 hours, and the extraction solution of Caesalpinia spinosaKuntze fruit compound is made；
95 mesh of extraction solution of Caesalpinia spinosaKuntze fruit compound is filtered, extract filtrate is made；
Extract filtrate is concentrated, extract concentrated solution is made；
By extract concentrated solution with 1:20 dipropylene glycol aqueous solution redissolves, and extract is made and redissolves liquid；
Extract is answered into solution sterilization, compound skin-whitening composition is made.
Embodiment 5
Climbing entada stem rattan is first taken, impurity is removed, sets in frying container, heated with slow boiling, stir-fry to color burn to yellow black,
When aromatic effusion, cool, used time crushing of drying in the air is taken out；Then it is carried out after the foreign matter in other above-mentioned three tastes herbal raw materials being chosen
Dries pulverizing processing, obtains herbal raw material powder；
By Caesalpinia spinosaKuntze fruit, short cut tobacco orchid root, the hard English walnut seeds of Bai Er, climbing entada stem rattan respectively press 50%, 24%, 14%,
Caesalpinia spinosaKuntze fruit compound is made in 12% ratio；
By Caesalpinia spinosaKuntze fruit compound and 90% ethanol solution by weight 1:40 are mixed, and mixture is made, by this
Mixture is heated to 82 DEG C, stirs heat reflux 3 hours, the extraction solution of Caesalpinia spinosaKuntze fruit compound is made；
85 mesh of extraction solution of Caesalpinia spinosaKuntze fruit compound is filtered, extract filtrate is made；
Extract filtrate is concentrated, extract concentrated solution is made；
By extract concentrated solution with 1:20 dipropylene glycol aqueous solution redissolves, and extract is made and redissolves liquid；
Extract is answered into solution sterilization, compound skin-whitening composition is made.
The formula of the present invention is not limited to embodiment 1-5, the study found that being combined using the others in the scope of the invention
With similar effect, for example, by Caesalpinia spinosaKuntze fruit, short cut tobacco orchid root, the hard English walnut seeds of Bai Er, climbing entada stem rattan respectively according to 34%,
20%, Caesalpinia spinosaKuntze fruit compound is made in 29%, 17% ratio；Or by Caesalpinia spinosaKuntze fruit, short cut tobacco orchid root, the hard English walnuts of Bai Er
Caesalpinia spinosaKuntze fruit compound is made in 43%, 22%, 16%, 19% ratio respectively in seed, climbing entada stem rattan；Final compound obtained
Lightening compositions can also play the effect of the present invention.
Embodiment 6：A kind of cleawhite over-night essences of the composition containing compound skin-whitening, includes each group of following mass percentage
Point, as shown in table 1-2：
Table 1
Table 2
| Project | Comparative example 1 | Comparative example 2 | Comparative example 3 | Comparative example 4 |
| Component | % | % | % | % |
| Compound skin-whitening composition | 5 | 5 | 5 | 5 |
| Glycerine | 10 | 10 | 10 | 10 |
| Glycerine polyacrylate | 0.5 | 0.5 | 0.5 | 0.5 |
| Sodium Hyaluronate | 0.05 | 0.05 | 0.05 | 0.05 |
| Urea | 1 | 1 | 1 | 1 |
| Anjidew NL50 | 0.5 | 0.5 | 0.5 | 0.5 |
| Polyquaternium -51 | 1 | 1 | 1 | 1 |
| Trehalose | 0.3 | 0.3 | 0.3 | 0.3 |
| Ethoxy urea | 0.5 | 0.5 | 0.5 | 0.5 |
| Ethyl alcohol | 2.5 | 2.5 | 2.5 | 2.5 |
| Serine | 0.5 | 0.5 | 0.5 | 0.5 |
| Sprout short stalk enzyme polysaccharide | 0.5 | 0.5 | 0.5 | 0.5 |
| Panthenol | 0.5 | 0.5 | 0.5 | 0.5 |
| Carbomer | 0.3 | 0.3 | 0.3 | 0.3 |
| Cremophor RH40 | 0.03 | 0.03 | 0.03 | 0.03 |
| Aminomethyl propanol | 0.3 | 0.3 | 0.3 | 0.3 |
| Potassium sorbate | 0.5 | 0.5 | 0.5 | 0.5 |
| Water | Surplus | Surplus | Surplus | Surplus |
Wherein, compound skin-whitening composition described in Essence 1-5 is respectively embodiment 1, embodiment 2, embodiment 3, implements
Compound skin-whitening composition described in example 4, embodiment 5；Preservative wherein described in Essence 1 is benzyl alcohol, institute in Essence 2
The preservative stated is potassium sorbate, and the preservative described in Essence 3 is potassium sorbate, and the preservative described in Essence 4 is
Benzyl carbinol, the preservative described in Essence 5 are potassium sorbate.
Wherein, the compound skin-whitening composition described in comparative example 1 by being prepared following preparation method：By Caesalpinia spinosaKuntze fruit and
75% acetone soln is by weight 1:25 are mixed, and mixture is made, which is heated to 85 DEG C, stir heat reflux
2 hours, the extraction solution of Caesalpinia spinosaKuntze fruit is made；90 mesh of extraction solution of Caesalpinia spinosaKuntze fruit is filtered, extract filtrate is made；It will carry
It takes object filtrate to concentrate, extract concentrated solution is made；By extract concentrated solution with 1:20 dipropylene glycol aqueous solution redissolves, and is made and carries
Object is taken to redissolve liquid；Extract is answered into solution sterilization, compound skin-whitening composition is made.
Wherein, the compound skin-whitening composition described in comparative example 2 by being prepared following preparation method：By short cut tobacco orchid root
Acetone soln with 75% is by weight 1:25 are mixed, and mixture is made, which are heated to 85 DEG C, stirring heat is returned
Stream 2 hours, is made the extraction solution of short cut tobacco orchid root；90 mesh of extraction solution of short cut tobacco orchid root is filtered, extract filter is made
Liquid；Extract filtrate is concentrated, extract concentrated solution is made；By extract concentrated solution with 1:20 dipropylene glycol aqueous solution is multiple
It is molten, extract is made and redissolves liquid；Extract is answered into solution sterilization, compound skin-whitening composition is made.
Wherein, the compound skin-whitening composition described in comparative example 3 by being prepared following preparation method：By the hard English walnuts of Bai Er
Seed and 75% acetone soln by weight 1:25 are mixed, and mixture is made, which are heated to 85 DEG C, stirring heat
Reflux 2 hours, is made the extraction solution of the hard English walnut seeds of Bai Er；90 mesh of extraction solution of the hard English walnut seeds of Bai Er is filtered, is made and carries
Take object filtrate；Extract filtrate is concentrated, extract concentrated solution is made；By extract concentrated solution with 1:20 dipropylene glycol is water-soluble
Liquid redissolves, and extract is made and redissolves liquid；Extract is answered into solution sterilization, compound skin-whitening composition is made.
Wherein, the compound skin-whitening composition described in comparative example 4 by being prepared following preparation method：First take climbing entada stem
Rattan removes impurity, sets in frying container, heated with slow boiling, fries to color burn to yellow black, and when aromatic effusion, taking-up is dried in the air
It is cool, then crush；By climbing entada stem rattan and 75% acetone soln by weight 1:25 are mixed, and mixture is made, by this
Mixture is heated to 85 DEG C, stirs heat reflux 2 hours, the extraction solution of climbing entada stem rattan is made；By the extraction of climbing entada stem rattan
90 mesh of solution filters, and extract filtrate is made；Extract filtrate is concentrated, extract concentrated solution is made；By extract concentrated solution
With 1:20 dipropylene glycol aqueous solution redissolves, and extract is made and redissolves liquid；Extract is answered into solution sterilization, compound skin-whitening group is made
Close object.
The effect of cleawhite over-night essences of present invention composition containing compound skin-whitening, is assessed
1, tyrosinase inhibition rate
Participating in B16 cell, there are mainly three types of enzymes：Tyrosinase, dopachrome interconvertible enzyme and dihydroxy indole carboxylic acid oxygen
Change enzyme.Tyrosinase category oxidoreducing enzyme, is the major rate-limiting enzyme of B16 cell, and tyrosinase can with its substrate l-tyrosine
Catalysis reaction occurs.
After being added to the reagent of tyrosinase activity inhibiting effect in experimental system, catalysis reaction can be generated
Inhibiting effect, by the light absorption value before and after measurement addition reagent at 475nm, to evaluate inhibition of the reagent to tyrosinase activity
Rate.Under different tested concentration, cleawhite over-night essences tyrosinase inhibition rate experimental result such as 3 institute of table of the composition containing compound skin-whitening
Show：
Table 3
As shown in Table 3, the cleawhite over-night essences of the composition containing compound skin-whitening has good tyrosinase inhibition rate, when tested
When a concentration of 1.0%, the cleawhite over-night essences tyrosinase inhibition rate of present invention composition containing compound skin-whitening reaches 70% or so, table
The cleawhite over-night essences of bright present invention composition containing compound skin-whitening has good tyrosinase inhibition rate；And in identical concentration
Under, the inhibiting rate of the cleawhite over-night essences tyrosinase of the composition containing compound skin-whitening is higher than the competing product of commercially available whitening.
2, melanocyte is tested
The Essence of composition containing compound skin-whitening is inquired into the proliferation of melanocyte and the influence of whitening function.Utilize B16
Melanoma cells measure influence of the Essence to melanocyte proliferation function, and to melanocyte synthesis of melanin
Influence.
(1) measurement (mtt assay) of cell proliferation rate
Murine melanoma B16 cells are cultivated, wait for that cell growth to logarithmic phase, is digested with 0.25% pancreatin, adjusts cell
A concentration of 5 × 104A/mL is inoculated in 96 orifice plates, per 100 μ L of hole.It is placed in incubator after cultivating for 24 hours, abandons supernatant, add
Add pastille culture medium, three concentration groups of each sample.Each concentration sets 3 multiple holes, and control group is not added with drug, instead accordingly
Solvent.After being incubated 72h, supernatant is abandoned after MTT solution 20 the μ L, 4h of 5g/L is added per hole, 100 μ of cell pyrolysis liquid is added per hole
L continues overnight incubation, each hole absorbance value, wavelength 570nm is detected in microplate reader.
Cell proliferation inhibition rate %=(1- medicine groups average absorbance value/control group average absorbance value) × 100
(2) measurement of B16 cell inhibiting rate
By B16 melanocytes with 20 × 104The density of a/mL is inoculated in 96 orifice plates.Per hole 1mL, after being incubated for 24 hours,
Supernatant is abandoned, adds drug to be measured (sample effect concentration is with aforementioned), after drug effect 72h, abandons supernatant, PBS washings, per hole
The 500 μ L of 1mo l/LNaOH solution containing 10%DMSO, concussion 5mi n is added.Measure each hole light absorption value under 490nm.
B16 cell inhibiting rate %=[1- (drug hole 490nm absorbance values/drug hole 570nm absorbances)/(control
Hole 490nm absorbance values/control wells 570nm absorbances)] × 100
After the cleawhite over-night essences tested material of various concentration composition containing compound skin-whitening is added, sample is to melanocyte black
Element synthesis inhibiting rate and cell proliferation inhibition rate experimental result are as shown in table 4：
The cleawhite over-night essences sample of 4 various concentration composition containing compound skin-whitening of table is to the proliferation inhibition rate of B16 cells and black
Pigment synthesis inhibiting rate
Conclusion：After the tested material of various concentration is added, the cleawhite over-night essences sample of the composition containing compound skin-whitening has preferably
Inhibition melanocyte synthesis of melanin effect, in tested concentration range (0.5-2.0%), B16 cell inhibiting rate
In 33-41%, show that the cleawhite over-night essences of the composition of the invention containing compound skin-whitening has good white-skinned face function, same concentrations item
Under part, the cleawhite over-night essences B16 cell inhibiting rate of the composition containing compound skin-whitening is higher than commercially available competing product, and cell proliferation
It influences to be higher than commercially available competing product.
3, crowd's investigation on probation
For the practical effect of test product, people has been carried out respectively to sample made from Essence 1-5 and comparative example 1-4
Group's investigation on probation distributes sample to the volunteer (wherein women 385) at 500 ages 28-42 Sui, health, life rule
Rule, no skin disease, volunteer are divided into 10 groups at random, and one of which is commercial product cleawhite over-night essences.Volunteer cleans face
It after skin, dries, appropriate Essence is applied to left side face with hand, right side face does not smear, as skin whitener test comparison
Area, tested period face do not smear other cosmetics or drug, and continuous use checks using effect in 30 days, and fills in investigation and ask
Volume.
It is classified the standard of judgement：
Significantly effectively：Opposite right side face, test block color spot problem are completely resolved, and skin smooth is for example new, the white and moist pool of jade-like stone.
Effectively：Opposite right side face, test block color spot is obviously thin out, forms color spot soon or even can completely disappear, skin
Skin becomes pale.
In vain：Opposite right side face, test block color spot do not change without significant change or intensification, the colour of skin.
Crowd tries out investigation result table 5
| Result on trial | Significantly effectively/example | Effectively/example | In vain/example |
| Essence 1 | 25 | 20 | 5 |
| Essence 2 | 23 | 23 | 4 |
| Essence 3 | 27 | 21 | 2 |
| Essence 4 | 41 | 9 | 0 |
| Essence 5 | 41 | 9 | 0 |
| Comparative example 1 | 1 | 8 | 41 |
| Comparative example 2 | 1 | 7 | 42 |
| Comparative example 3 | 0 | 6 | 44 |
| Comparative example 4 | 1 | 5 | 44 |
| Commercial product | 12 | 20 | 18 |
Conclusion：In the cleawhite over-night essences sample of the composition containing compound skin-whitening, it is (Caesalpinia spinosaKuntze fruit, short to be added to whole components
The hard English walnut seed of cut tobacco orchid root, Bai Er, climbing entada stem rattan) Essence 1-5 effects (significantly effectively >=23) it is more independent than only adding
The effect (significantly effectively≤1) of the comparative example 1-4 of ingredient will be got well, and only the effect of the comparative example 1-4 of addition separate constituent is simultaneously
Not significantly, the results showed that each ingredient has mutually association in skin whitening and dispelling stain etc. in the compound skin-whitening composition
Effect together simultaneously obtains preferable effect.
By Essence 1-5 with commercial product (significantly effective 12) comparison it is found that the whitening effect of the present invention is more preferable.
It is learnt through questionnaire, makes have essence liquid product of the present invention, mild non-stimulated, skin sense is fine.
Embodiment described above is only that the preferred embodiment of the present invention is described, not to the model of the present invention
It encloses and is defined, under the premise of not departing from design spirit of the present invention, technical side of the those of ordinary skill in the art to the present invention
The various modifications and improvement that case is made should all be fallen into the protection domain of claims of the present invention determination.
Claims (10)
1. a kind of preparation method of compound skin-whitening composition, which is characterized in that include the following steps：
(1) prepare herbal raw material in proportion：The herbal raw material is made of major ingredient and auxiliary material, and the major ingredient is Caesalpinia spinosaKuntze
Fruit, weight percent 30-50%, auxiliary material are short cut tobacco orchid root, the hard English walnut seeds of Bai Er, climbing entada stem rattan, weight percent score
It Wei not 20-30%, 10-30% and 10-20%；
(2) it pre-processes：Climbing entada stem rattan is first taken, impurity is removed, sets in frying container, heated with slow boiling, is fried to color burn extremely
Yellow black, when aromatic effusion, taking-up is dried in the air cool, and the used time crushes；Then the foreign matter in other above-mentioned three tastes herbal raw materials is chosen
Dries pulverizing processing is carried out after going out, and obtains herbal raw material powder；Four kinds of herbal powders are uniformly mixed again, Caesalpinia spinosaKuntze fruit is made
Compound；
(3) it extracts：The Caesalpinia spinosaKuntze fruit compound and solvent are mixed and extracted, extraction solution is obtained；
(4) it filters：The extraction solution is filtered, extract filtrate is obtained；
(5) it concentrates：The extract filtrate is concentrated, extract concentrated solution is obtained；
(6) it redissolves：The extract concentrated solution is redissolved, extract is obtained and redissolves object；
(7) it sterilizes, the extract is redissolved into object sterilizing, obtains compound skin-whitening composition.
2. the preparation method of compound skin-whitening composition according to claim 1, which is characterized in that step is pierced described in (3)
Mysorethorn fruit compound and solvent are by weight 1:（10-40）It is mixed, extracts 1-3 hours, must carry at a temperature of 80-90 DEG C
Take solution.
3. the preparation method of compound skin-whitening composition according to claim 1, which is characterized in that mistake described in step (4)
Filter is the filtering of 80-100 mesh.
4. the preparation method of compound skin-whitening composition according to claim 1, which is characterized in that multiple described in step (6)
It is molten to use 1:20 dipropylene glycol aqueous solution redissolves.
5. the preparation method of compound skin-whitening composition according to claim 1, which is characterized in that molten described in step (3)
Agent is the ethanol solution of 60-90% or the acetone soln of 60-90%.
6. the preparation method of compound skin-whitening composition according to claim 1, which is characterized in that step carries described in (3)
It is taken as being stirred at reflux extraction or ultrasonic extraction.
7. application of the compound skin-whitening composition described in any one of claim 1-6 claims in cosmetics.
8. a kind of cosmetics, which is characterized in that including the compound skin-whitening group described in any one of claim 1-6 claims
Object is closed, the dosage of the compound skin-whitening composition is the 1-10% of final products weight；Preferably, the compound skin-whitening composition
Dosage be final products weight 2-8%.
9. cosmetics according to claim 8, which is characterized in that further include solvent, emollient, solubilising in the cosmetics
Agent, antioxidant, pH adjusting agent, penetrating agent, emulsion stabilizer, preservative, moisturizer, thickener, neutralizer, chelating agent, table
One kind in face activating agent, emulsifier, essence, colorant, sun-screening agent, skin conditioning agent, active matter and functional additive or
It is several.
10. cosmetics according to claim 8, which is characterized in that the cosmetics be toner, shower cream, face cleaning cream,
Facial mask, face cream, gel, lotion, Essence, makeup remover or transparent soap.
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