CN103876989A - Application of tree peony bark extract to preparation of skin whitening composition for external use and preparation method of tree peony bark extract - Google Patents
Application of tree peony bark extract to preparation of skin whitening composition for external use and preparation method of tree peony bark extract Download PDFInfo
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Abstract
The invention relates to an application of a tree peony bark extract to preparation of a skin whitening composition for external use and a preparation method of the tree peony bark extract. The tree peony bark extract contains a component for inhibiting tyrosinase. The preparation method of the tree peony bark extract comprises the steps of extracting tree peony barks by using an organic alcohol water solution to obtain an extracting solution; concentrating the extracting solution, and drying to obtain a crude extract; dispersing the crude extract by using water to form a suspension liquid, separating and purifying the suspension liquid by using macroporous resin, and eluting many times to obtain an eluant; concentrating the eluant, and drying to obtain the tree peony bark extract. The tree peony bark extract obtained by using the preparation method provided by the invention plays a skin whitening role through inhibiting the activity of tyrosinase and further inhibiting the generation of melanin, also has certain stability and can be used for preparing a composition for external use.
Description
Technical field
The present invention relates to the purposes of Cortex Moutan (Paeoniasuffruticosa) extract, especially it is for the purposes of skin-whitening, the preparation method of this Cortex Moutan extract, and comprise the lightening compositions of this Cortex Moutan extract.
Background technology
Melanocyte is a kind of biochrome of high molecular, can play the effect of shielding of ultraviolet, thereby has the function of protecting skin from ultraviolet.At present known has three kinds, is respectively eumelanin (Eumelanin), pheomelanin (Pheomelanin) and neuromelanin (Neuromelanin), and the melanocyte of indication is eumelanin conventionally.After skin is by ultraviolet radiation, in body, can produce a large amount of melanin with protection skin.Melanic content is the important factor in order of the skin color depth.Melanic content is more, and the colour of skin of skin is just more black.Thereby the material that can play check melanin generation is regarded as having the effect of skin whitening.
Tryrosinase is a kind of polyphenol oxidase, belongs to redox enzymes, and it is the main rate-limiting enzyme in melanin forming process, and the size of this enzymatic activity has determined the quantity that melanin forms.Tyrosine in Skin Cell, under the effect of tryrosinase, is subject to the impact of various factors in environment, is oxidized to gradually DOPA and DOPA quinone, and DOPA quinone passes through a series of metabolic response again, is polymerized to 5,6-istain, is finally progressively converted into dermal melanin.
Cortex Moutan is the dry root bark of ranunculaceae peony (PaeoniasuffruticosaAndr), there is the effect of clearing away heat and cooling blood, blood circulation promoting and blood stasis dispelling, mainly contain maculae caused by violent heat pathogen, haematemesis, night fever abating at dawn, lossless hectic fever due to YIN-deficiency, amenorrhea dysmenorrhea, carbuncle sore tumefacting virus, fall and flutter the effects such as grieved.And according to document and related data, that Cortex Moutan has is anticancer, antianaphylaxis asthma and the pharmacological action that protects the liver, its main component paeonol more can reduce the neurotoxicity of D-galactosamine induction aged mouse, and can improve its cognitive function.But the application of Cortex Moutan crude extract in skin-lightening cosmetic but has no report.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of new purposes of Cortex Moutan extract.
Another technical problem to be solved by this invention is to provide the preparation method of Cortex Moutan extract in such use.
A technical problem more to be solved by this invention is to provide a kind of lightening compositions.
The present invention solves the problems of the technologies described above taked technical scheme and is to provide the purposes of a kind of Cortex Moutan extract for the preparation of external preparation for skin lightening compositions.
Term used herein " skin " not only comprises and covers the face of animal or the tissue of body surface, but also comprises scalp and hair.
Here the term " extract " that used refers to, extracts composition and any material of obtaining by any method from natural material.Described extract comprises, from natural materials with an organic solvent or the composition that dissolves in described solvent of water extraction, and the special component extracting from natural materials.
Described Cortex Moutan extract specifically comprises the composition for restraint of tyrosinase, and its activity to tryrosinase has significant inhibition.
The present invention solves the problems of the technologies described above another taked technical scheme and is to provide the preparation method of Cortex Moutan extract in such use, and the preparation of described Cortex Moutan extract comprises the steps:
(1) Cortex Moutan is used to Organic Alcohol extraction with aqueous solution, obtained extracting solution;
(2) extracting solution is concentrated, dry, obtains crude extract extractum;
(3) crude extract extractum water is dispersed into suspension, carries out separation and purification by macroporous resin column, and eluting repeatedly, obtains eluent;
(4) eluent is concentrated, dry, obtains Cortex Moutan extract.
The concentrated mode of extracting solution is evaporation and concentration or concentrating under reduced pressure in step (2), and described drying mode is drying under reduced pressure or vacuum lyophilization, volatilizees completely to solvent.
The concentrated mode of eluent is evaporation and concentration or concentrating under reduced pressure in step (4), and described drying mode is drying under reduced pressure or vacuum lyophilization, volatilizees completely to solvent.
Described Cortex Moutan is the dry root bark of ranunculaceae peony.
In step (1), add 70~95% Organic Alcohol aqueous solution of 10~30 times of weight of Cortex Moutan, reflux, extract, 1~10 hour at 65~95 ℃.
In step (1), be preferably 15~25 times of Cortex Moutan weight for the Organic Alcohol aqueous solution extracting, more preferably 20 times.Extract preferably 70~90 ℃ of temperature, preferably 3~7 hours extraction time, more preferably 4~6 hours.
In step (3), the Organic Alcohol aqueous solution eluting that the volume fraction of the Organic Alcohol aqueous solution eluting that the volume fraction of the Organic Alcohol aqueous solution eluting that is 20%~40% by the volume fraction of the water elution of 2~5 times of resin column volumes, 1~3 times of resin column volume successively, 2~4 times of resin column volumes is 45%~70%, 2~5 times of resin column volumes is 85%~95%; Described Organic Alcohol is one or more in methanol, ethanol and propanol.
The ethanol water eluting that the volume fraction of the ethanol water eluting that elution step is preferably 25~35% by the volume fraction of the water elution of 3~4 times of resin column volumes, 2~3 times of resin column volumes successively, ethanol water eluting that the volume fraction of 2.5~3.5 times of resin column volumes is 55~65% and 2.5~3.5 times of resin column volumes is 88~92%.After eluting, through concentrated and dry, obtain the Cortex Moutan extract at each position.
The present invention solves the problems of the technologies described above a taked technical scheme again and is to provide a kind of external preparation for skin lightening compositions, comprises the Cortex Moutan extract being obtained by above-mentioned preparation method.
The mass percent of described Cortex Moutan extract in total composition is 0.01~10%.
The mass percent of Cortex Moutan extract in total composition is for being preferably 0.05 ~ 2%.
Described topical composition is cosmetics for external use compositions or topical drug compositions.
Described compositions also comprises one or more in whitening agent except Cortex Moutan extract, defying age composition, antioxidant, vitamin, polypeptide, polysaccharide, sphingolipid.Do not affecting under the prerequisite of object of the present invention and effect, those skilled in the art can easily select the content of the above-mentioned constituent in described cosmetic composition.
Described compositions can also comprise wetting agent, emollient, surfactant, ultraviolet absorber, antiseptic, disinfectant, antioxidant, pH adjusting agent, organic and/or inorganic pigment, flavoring agent, coolant, antiperspirant etc. and can improve composition main effect and that do not affect its main effect of described compositions.
The dosage form of described compositions is solution, emulsion, suspension, cream, gel, foam or spraying.
For cosmetics for external use compositions, can be for being suitable for any dosage form of partial smearing.For example, described cosmetic composition can be the compositions of solid, solution, emulsion (emulsion oil-in-water, water-in-oil emulsion), suspension, cream, gel, paste, foam or Sprayable, but is not limited to these dosage forms.These dosage forms can be prepared by any common method known in the art.
For externally-applied medicinal composition, can also be for being suitable for any dosage form of partial smearing.For example, described pharmaceutical composition can be the compositions of solid, solution, emulsion (emulsion oil-in-water, water-in-oil emulsion), suspension, cream, gel, paste or Sprayable, but is not limited to these dosage forms.
According to any common method known in the art, can by with surfactant, excipient, wetting agent, emulsifying agent, suspending agent, prepare the dosage form of described pharmaceutical composition for the salt of osmotic pressure or buffer agent, coloring agent, flavoring agent, stabilizing agent, antiseptic or other conventional additives.
Beneficial effect of the present invention:
The Cortex Moutan extract being obtained by preparation method of the present invention, the generation that reaches check melanin (melanin) by the activity of restraint of tyrosinase (tyrosinase) shows skin-whitening effect, and there is certain stability, can, for the preparation of composition for external application, demonstrate skin whitening efficacy.
The specific embodiment
To further describe the present invention by describing embodiment below, but content of the present invention is not limited only to this.
Embodiment 1: the preparation of Cortex Moutan extract
Get Cortex Moutan and clean, be dried, remove impurity, for subsequent use.
(1) take a certain amount of Cortex Moutan, extract with 20 times of amount ethanol waters of 90%, at 70~90 ℃, extract 4~6 hours, extracting solution volatilizees completely through leaving standstill, filter, concentrate, be dried to solvent, obtains Cortex Moutan crude extract extractum (extract A);
(2) extractum is with after aqueous dispersion, adopt macroporous adsorptive resins to carry out separation and purification, the ethanol water that the volume fraction of the ethanol water that is 30% by the volume fraction of the water of 4 times of resin column volumes, 3 times of resin column volumes successively, ethanol water that the volume fraction of 3 times of resin column volumes is 60% and 3 times of resin column volumes is 90% carries out eluting, collect respectively the eluent at each position, concentrated, vacuum drying or lyophilization to solvent volatilizees completely, obtains the Cortex Moutan extract at each position.Wherein, wash the eluent obtaining with water and be designated as extract B, be designated as extract C with the eluent that 30% ethanol water eluting obtains, be designated as extract D with the eluent that 60% ethanol water eluting obtains, be designated as extract E with the eluent that 90% ethanol water eluting obtains.
the toxicity assessment of Cortex Moutan crude extract extractum (extract A) in B16 cell
Get the cell monolayer of trypsinization logarithmic (log) phase, and by cell harvesting to culture bottle; Count and adjust concentration of cell suspension to 2.5 × 10
3~50 × 10
3individual/mL; Spread 96 orifice plates and be placed in 5%CO
2in incubator, cultivate 24h; Add Cortex Moutan extract A sample or control sample continuation cultivation 48h with the variable concentrations of dissolution with solvents; Take out, add the MTT(tetrazolium bromide of the 5mg/mL now joining) solution, every hole 20 μ L also continue to cultivate 4h; Abandon the dimethyl sulfoxine that supernatant every hole add 150 μ L, place 15 minutes, dissolve bluish violet crystallization; Shake and use microplate reader to test absorbance under 490nm, and calculating the cell survival rate of each medicine under selected concentration.(survival rate=
× 100%)
The cytotoxicity result of extract A is as shown in table 1.
As shown in Table 1, the Cortex Moutan extract A being obtained by step (1) is under 0.08,0.4,2,10 and 50 μ g/mL, to B16 cell no cytotoxicity.Cell survival rate exceedes this extract A of 100% expression, in the time of finite concentration, B16 cell is had to certain proliferation function, therefore can regard it as and not have cytotoxicity.
cortex Moutan crude extract extractum (extract A) suppresses the evaluation of B16 cell tyrosinase activity
Get the cell monolayer of trypsinization logarithmic (log) phase, by cell harvesting to culture bottle; Count and adjust concentration of cell suspension to 2.5 × 10
3~50 × 10
3individual/mL; Bed board is also placed in containing 5%CO
2in incubator, cultivate 24h; Add Cortex Moutan extract A sample or the control sample of variable concentrations and continue to cultivate 48h; Take out, abandon supernatant and add the 0.5% deoxycholic acid sodium solution of now joining, every hole 100 μ L; Shake and add the 0.1%L-Dopa(levodopa of now joining), every hole 10 μ L; In water-bath, after water-bath 30min, taking-up is measured absorbance by microplate reader under 475nm, and calculates each medicine tyrosinase inhibition rate to B16 cell under selected concentration.(suppression ratio=1-
× 100%)
Extract A is as shown in table 2 to tyrosinase activity suppression ratio result.
As shown in Table 2, the Cortex Moutan extract A being obtained by step (1) shows certain tyrosinase inhibitory action, therefore can do further separation, to obtain active better Cortex Moutan extract.
cortex Moutan extract B, C, D and the E toxicity assessment in B16 cell
According to the toxicity assessment method of extract A before, Cortex Moutan extract B, C, D and E to be evaluated B16 cytotoxicity size, result is as shown in table 3.
As shown in Table 3, the each position Cortex Moutan extract B, C, D and the E that are obtained by step (2) are under 0.25,2.5 and 25 μ g/mL, to B16 cell no cytotoxicity.
cortex Moutan extract B, C, D and E suppress the evaluation of B16 cell tyrosinase activity
According to the inhibition B16 cell tyrosinase activity evaluation methodology of extract A before, Cortex Moutan extract B, C, D and E to be evaluated for B16 cell tyrosinase activity suppression ratio size, result is as shown in table 4.
As shown in Table 4, the each position Cortex Moutan extract B, C, D and the E that are obtained by step (2) show certain B16 cell tyrosinase inhibitory action, wherein, under same concentrations (25 μ g/mL), extract D is the strongest to the inhibition of B16 cell tyrosinase activity.
cortex Moutan extract B, C, D and E suppress the evaluation of Mushroom Tyrosinase activity
Preparation pH=6.8, the phosphate buffered solution of 0.1M.Get 885 μ LPBS, successively adding 15 μ L concentration is extract B, extract C, extract D or the extract E of 150 μ g/mL, and 100 μ L(100U) Mushroom Tyrosinase solution, mix.30 ℃ of water-baths 10 minutes, then add 80 μ L, 0.1% L-Dopa(levodopa) solution, water-bath, after 20 minutes, is measured absorbance under 475nm.
Extract B, C, D and E are as shown in table 5 to tyrosinase activity suppression ratio result.
As shown in Table 5, the each position Cortex Moutan extract B, C, D and the E that are obtained by step (2) show certain Mushroom Tyrosinase inhibitory action, and under same concentrations, extract D is the strongest to the inhibition of Mushroom Tyrosinase activity.
Embodiment 2: the preparation of Cortex Moutan extract
Get Cortex Moutan and clean, be dried, remove impurity, for subsequent use.
(1) take a certain amount of Cortex Moutan, extract with 25 times of amount ethanol waters of 80%, extract 5~7 hours at 80~90 ℃, extracting solution volatilizees completely through leaving standstill, filter, concentrate, be dried to solvent, obtains Cortex Moutan crude extract extractum;
(2) extractum is with after aqueous dispersion, adopt macroporous adsorptive resins to carry out separation and purification, the ethanol water eluting that the volume fraction of the ethanol water that is 35% by the volume fraction of the water of 3 times of resin column volumes, 2 times of resin column volumes successively, ethanol water that the volume fraction of 3.5 times of resin column volumes is 55% and 2.5 times of resin column volumes is 92%, collect respectively the eluent at each position, concentrated, vacuum drying or lyophilization to solvent volatilizees completely, obtains the Cortex Moutan extract at each position.
Embodiment 3: the preparation of Cortex Moutan extract
Get Cortex Moutan and clean, be dried, remove impurity, for subsequent use.
(1) take a certain amount of Cortex Moutan, extract with 15 times of amount ethanol waters of 95%, at 70~80 ℃, extract 3~5 hours, extracting solution volatilizees completely through leaving standstill, filter, concentrate, be dried to solvent, obtains Cortex Moutan crude extract extractum (extract A);
(2) extractum is with after aqueous dispersion, adopt macroporous adsorptive resins to carry out separation and purification, the ethanol water eluting that the volume fraction of the ethanol water that is 25% by the volume fraction of the water of 3.5 times of resin column volumes, 2.5 times of resin column volumes successively, ethanol water that the volume fraction of 2.5 times of resin column volumes is 65% and 3.5 times of resin column volumes is 88%, collect respectively the eluent at each position, concentrated, vacuum drying or lyophilization to solvent volatilizees completely, obtains the Cortex Moutan extract at each position.
The whitening emulsion that application examples 1 comprises Cortex Moutan extract
By the Cortex Moutan extract D making in embodiment 1, prepare lightening compositions (emulsion) by the proportioning in table 6.
assessment containing the lightening compositions of Cortex Moutan extract to cutaneous safety
To 19 adult females and 11 adult males carry out paster test (mean age: 33), to detect the compositions of above-described embodiment to the safety of skin.
Tested material is put into speckle examination device, and consumption is about 0.020~0.025g(solid or semisolid) or 0.020~0.025mL(liquid, can drip on the appended filter paper of speckle examination device and be placed in speckle examination device).When tested material is the cosmetics finished product original, control wells is blank (declining any material), when tested material is the cosmetics after dilution, uses the diluent of these cosmetics in control wells.The non-stimulated tape sticker of speckle examination device that is added with tested material is spread on to experimenter's back or the bent side of forearm, gently press and make it to stick equably on skin with palm, continue 24h.
30min after removal tested material speckle examination device observes dermoreaction after impression disappears.As negative in result, after patch test, 24h and 48h observe once respectively again.By table 7(skin adverse reaction grade scale table) record reaction result.
As shown in the result of above-mentioned table 7, lightening compositions is Diazolidinyl Urea not.Therefore, can show that cosmetic composition of the present invention is safe to skin.
contain the skin whitening efficacy test of the lightening compositions of Cortex Moutan extract
The whitening emulsion preparing is used in crowd, choose 30 adult females of 18~40 years old, 33 years old mean age.Skin belongs to normal condition, and health is normal, and unglazed allergies, without cosmetic allergic contact dermatitis history.Adopt double-blind placebo-controlled contrast principle, lightening compositions (containing the Cortex Moutan extract D by 60% ethanol water eluting) and matched group (not containing Cortex Moutan extract) are evenly spread upon respectively on the forearm of two arms, every arm is smeared a kind of sample, and every day sooner or later respectively once.Measuring the instrument of melanin content is the MPA580 that is furnished with MX18 probe, and inside tester's forearm, same position is chosen the test zone that a radius is 2cm, respectively before use with 4th week and the 8th week melanic content of mensuration, test result is as shown in table 8.
As shown in Table 8, compared with matched group, smeared the skin of application examples 1 whitening emulsion, melanin content significantly reduces in the time of 4 weeks and 8 weeks, and from visually observing, compared with matched group, skin has more reflecting feel.Prove that application examples 1 whitening emulsion has whitening effect.
The cleawhite pack that application examples 2 comprises Cortex Moutan extract
Prepare facial film according to the component in existing method use table 9.
The whitening gelatin that application examples 3 comprises Cortex Moutan extract
Prepare gel according to the component in existing method use table 10.
The whitening water that application examples 4 comprises Cortex Moutan extract
Prepare water according to the component in existing method use table 11.
The whitening emulsion that application examples 5 comprises Cortex Moutan extract
Prepare emulsion according to the component in existing method use table 12.
The whitening ointment that application examples 6 comprises Cortex Moutan extract
Prepare ointment according to the component in existing method use table 13.
Claims (10)
1. Cortex Moutan extract is for the preparation of the purposes of external preparation for skin lightening compositions.
2. Cortex Moutan extract according to claim 1, for the preparation of the purposes of external preparation for skin lightening compositions, is characterized in that: the composition that described Cortex Moutan extract comprises restraint of tyrosinase.
3. according to the preparation method of Cortex Moutan extract in purposes described in claim 1 or 2, it is characterized in that: comprise the steps:
(1) Cortex Moutan is used to Organic Alcohol extraction with aqueous solution, obtained extracting solution;
(2) extracting solution is concentrated, dry, obtains crude extract extractum;
(3) crude extract extractum water is dispersed into suspension, carries out separation and purification by macroporous resin column, and eluting repeatedly, obtains eluent;
(4) eluent is concentrated, dry, obtains Cortex Moutan extract.
4. the preparation method of Cortex Moutan extract according to claim 3, is characterized in that: described Cortex Moutan is the dry root bark of ranunculaceae peony.
5. the preparation method of Cortex Moutan extract according to claim 3, is characterized in that: in step (1), add 70~95% Organic Alcohol aqueous solution of 10~30 times of weight of Cortex Moutan, reflux, extract, 1~10 hour at 65~95 ℃; In step (3), the Organic Alcohol aqueous solution eluting that the volume fraction of the Organic Alcohol aqueous solution eluting that the volume fraction of the Organic Alcohol aqueous solution eluting that is 20%~40% by the volume fraction of the water elution of 2~5 times of resin column volumes, 1~3 times of resin column volume successively, 2~4 times of resin column volumes is 45%~70%, 2~5 times of resin column volumes is 85%~95%, obtains the eluent at each position; Described Organic Alcohol is one or more in methanol, ethanol and propanol.
6. an external preparation for skin lightening compositions, comprises the Cortex Moutan extract being obtained by the preparation method of one of claim 3 to 5.
7. external preparation for skin lightening compositions according to claim 6, is characterized in that: described Cortex Moutan extract is the extract being obtained by 45%~70% Organic Alcohol aqueous solution eluting.
8. according to the external preparation for skin lightening compositions described in claim 6 or 7, it is characterized in that: the mass percent of described Cortex Moutan extract in total composition is 0.01~10%, is preferably 0.05 ~ 2%.
9. according to the external preparation for skin lightening compositions described in claim 6 or 7, it is characterized in that: described compositions also comprises one or more in whitening agent except Cortex Moutan extract, defying age composition, antioxidant, vitamin, polypeptide, polysaccharide, sphingolipid.
10. according to the external preparation for skin lightening compositions described in claim 6 or 7, it is characterized in that: the dosage form of described compositions is solution, emulsion, suspension, cream, gel, foam or spraying.
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CN104127353A (en) * | 2014-07-31 | 2014-11-05 | 广州丹奇日用化工厂有限公司 | Cosmetic composition and preparation method thereof |
CN104288094A (en) * | 2014-09-30 | 2015-01-21 | 江苏奇力康皮肤药业有限公司 | Method for preparing paeonol ointment for anti-inflammation and anti-pruritus of skin |
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