CN102362842B - Preparation and application of traditional Chinese medicine compound extract with whitening and freckle removing effects - Google Patents
Preparation and application of traditional Chinese medicine compound extract with whitening and freckle removing effects Download PDFInfo
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Abstract
The invention relates to a traditional Chinese medicine compound extract with whitening and beverage removing effects. The traditional Chinese medicine compound extract is an extract prepared by using ten kinds of traditional Chinese medicine such as patrinia villosa juss, shinyleaf yellowhorn kernels, cortex dictamni, hairyvein agrimony, rhodiola root, cassia seeds, semen boitae, golden buckwheat rhizome, peppermint, liquorice and the like as raw materials and adopting the carbon dioxide supercritical extraction technology and the macroporous absorbent resin enrichment technology at certain technological parameters. The extract has an obvious tyrosinase activity inhibition effect and a melanin generation inhibition effect and can be directly and safely applied to the preparation of cosmetics and external use medicine preparations with the whitening and beverage removing effects.
Description
Technical field
The present invention relates to a kind of Chinese medicine compound extract, specifically relate to a kind of employing carbon dioxide supercritical fluid extraction technology and macroporous adsorbent resin beneficiation technologies, be the extract that raw material makes with patrima villosa grass, Lignum Xanthoceratis kernel, Cortex Dictamni, Herba Agrimoniae, Radix Rhodiolae, Semen Cassiae, Semen Platycladi, Rhizoma Fagopyri Dibotryis, Herba Menthae, Radix Glycyrrhizae, this extract has tangible tyrosinase activity inhibitory action and melanin generates inhibitory action.The invention still further relates to the application of this extract in pharmaceutical preparation, health food and the cosmetics of the class of dispelling are whitened in preparation.
Background technology
Loving beauty is part of human nature.Along with growth in the living standard, people are more and more higher to the requirement of quality of life, more and more pay attention to whitening naturally of skin, and facial area seem particularly important.Chloasma is a kind of acquired pigmentation dermatosis, is to be changed by the limitation due to the multiple reason, scrambling skin pigment, and clinical manifestation is the symmetry pigmentation, be the butterfly aliform, the lighter is yellowish or light brown, and the some lamellar intersperses among the buccal both sides, and is multiple with the outside under the eye; Weight person is brown deeply or light/dark balance is distributed in face.Primary disease is mainly in young and middle-aged women, does not generally have any subjective symptoms, course of disease stubbornness, and it is attractive in appearance to have a strong impact on the patient, brings many worries and misery for patient's spirit and life aspect.
Chloasma pathogeny complexity, melanocyte too much are its core pathological manifestations.Its definite pathogenesis is not bright so far, it is generally acknowledged all multifactor relevantly with melanocyte Developmental and Metabolic Disorder, endocrine disturbance, gestation, oral contraceptive, inherited genetic factors, oxygen-derived free radicals, ultraviolet radiation, tryrosinase dysfunction etc., wherein the skin melanocyte Developmental and Metabolic Disorder that causes such as endocrine disturbance, heredity, ultraviolet radiation is the main cause of its morbidity.
Modern medicine treatment chloasma mainly comprises uses lucifuge agent (as zinc oxide, titanium dioxide etc.), and system uses vitamins, tranamic acid, melatonin etc.; But substantially based on the local topical chemical decolorization, as sterin in hydrogen ketone, Azelaic Acid, kojic acid, glutathion, indometacin, retinoic acid, phenolic compound, the cortex etc.; Also having chemistry to strip off horny layer or chemical skin sand grinds off except melanocyte and with dye laser and destroys melanin granule etc., most methods is because causing inhomogeneous pigment and take off toxic and side effects such as mistake, telangiectasis, contact dermatitis or itself is unstable, slow curative effect easily recurs, and the fraud that causes permanent decolouring and medicine source property moth patch is arranged, and fails to apply.
Chinese medicine is called chloasma " chloasma ", " dusty face ", " chloasma hepaticum " etc., the Golden Mirror of Medicine cloud: " dim for a long time idle dark as dirt, former form in the troubled thoughts depression ".To the symptom of primary disease cloud again: " first improvement such as dust and dirt, with the passing of time black sapropelitic shape is withered secretly not damp, not of uniform size.Little person such as foxtail millet grain Semen Phaseoli, big person is like Semen Nelumbinis or long or oblique or circle.Equal with skin, the women has it more ".Think the liver,kidney,spleen functional disorder, QI-blood circulation is not normal, makes the venation inanition can not go up China in face, or the turbid resistance network of the stasis of blood, and penting up skin is the main cause of chloasma morbidity.
The Chinese medicine chloasma is stressed internal organs negative and positive of qi and blood balance, pays attention to regulating functions of ZANG FU-organs, controls with blood circulation promoting and blood stasis dispelling dispersing the stagnated live-QI to relieve the stagnation of QI, nourishing the liver and kidney, strengthening spleen and nourishing blood.Or treatment by oral administration of medicines is main or external treatment is main, or interior external treatment and usefulness.The treatment by Chinese herbs chloasma is with a long history, in allusions such as Shennong's Herbal, " Mingyi Bielu ", Holy Benevolent Prescriptions, " prescriptions worth thousand gold " doctors nationality, just clearly put down in writing can " dark complexion of dispelling, good color " square medicine.A large amount of clinical practices prove that the Chinese medicine chloasma demonstrates certain advantage, have that outstanding integrally-regulated, stable curative effect, side effect are little, the patient is easy to advantages such as acceptance, and curative effect is than good with western medicine merely.But the treatment by Chinese herbs chloasma exists still that slow curative effect, cure rate are not high, the course for the treatment of is long, take shortcoming such as Chinese medicine inconvenience for a long time, has influenced the compliance for the treatment of.
In a word, medicine and the means of integrative therapy chloasma are many, but or because of curative effect too slowly or too many because of untoward reaction, do not have always a kind of evident in efficacy, safely, the therapeutic scheme of generally acknowledging, make doctors and patients all to be satisfied with, this needs the utmost point not conform to clinical practice.Therefore, seek the Chinese prescription of reasonable recipe, determined curative effect, and be prepared into efficient, safe Chinese medicine whitening and speckle dispelling extract by the modern Chinese medicine extractive technique, become a research focus of present Chinese medicine beauty treatment fields.
Summary of the invention
An object of the present invention is to provide a kind of tyrosinase activity inhibitory action and melanin with excellence and generate inhibiting Chinese medicine compound extract.
Another object of the present invention is to propose the application of this Chinese medicine compound extract in pharmaceutical preparation, health food and the cosmetics of preparation whitening and speckle dispelling.
In order to achieve the above object, the present invention has taked following technical scheme:
Prescription medical material and ratio are: in parts by weight, and 5 parts of patrima villosa grass, 5 parts of Lignum Xanthoceratis kernel, 3 parts of Cortex Dictamni, 3 parts of Herba Agrimoniaes, 3 parts of Radix Rhodiolaes, 3 parts of Semen Cassiaes, 3 parts of Semen Platycladi, 3 parts of Rhizoma Fagopyri Dibotryiss, 2 parts of Herba Menthaes, 1 part in Radix Glycyrrhizae.
In parts by weight, 5 parts of patrima villosa grass, 5 parts of Lignum Xanthoceratis kernel, 3 parts of Cortex Dictamni, 3 parts of Herba Agrimoniaes, 3 parts of Radix Rhodiolaes, 3 parts of Semen Cassiaes, 3 parts of Semen Platycladi, 3 parts of Rhizoma Fagopyri Dibotryiss, 2 parts of Herba Menthaes, 1 part in Radix Glycyrrhizae, it is an amount of to take by weighing each medical material, mix and pulverize, cross No. 2 sieves of pharmacopeia, medicated powder is packed in the carbon dioxide supercritical fluid extraction device, add the entrainer dehydrated alcohol, consumption is 0.5ml/g medicated powder, and setting extraction temperature is 45 ℃, and extracting pressure is 40MPa, resolving I pressure is 10MPa, temperature is 40 ℃, resolving II pressure is 4MPa, temperature is 25 ℃, and the extraction time is 2h, gets pale brown color thickness grease.With the medicinal residues behind the previous step carbon dioxide supercritical fluid extraction, added 10 times of amount 70% soak with ethanol 12 hours, heating and refluxing extraction 2 hours, filter, in medicinal residues, add 5 times of amount 70% alcohol heating reflux extraction 2 hours again, filter, merging filtrate, drying under reduced pressure gets extractum.Extractum is added suitable quantity of water dilution back to be mixed with isopyknic D-101 type macroporous adsorbent resin and mixes thoroughly, be splined in the D-101 type macroporous adsorptive resins (applied sample amount is calculated as 1: 2 with the ratio of crude drug amount and amount of resin), static adsorption 30 minutes, carry out eluting with 20% ethanol of the water of 10 column volumes, 10 column volumes and 50% ethanol of 10 column volumes successively then, collect 50% ethanol elution, pulverize behind the drying under reduced pressure, make brown ceramic powder.The pale brown color thickness grease that this powder and carbon dioxide supercritical fluid extraction are made mixes, and namely gets Chinese medicine compound extract of the present invention.
As to improvement of the technical scheme with replenish:
1, the relative quantity of prescription Chinese crude drug can be adjusted according to the ratio of: patrima villosa grass 4-6 part, Lignum Xanthoceratis kernel 4-6 part, Cortex Dictamni 2-4 part, Herba Agrimoniae 2-4 part, Radix Rhodiolae 2-4 part, Semen Cassiae 2-4 part, Semen Platycladi 2-4 part, Rhizoma Fagopyri Dibotryis 2-4 part, Herba Menthae 1-3 part, Radix Glycyrrhizae 1-2 part.
2, when carrying out carbon dioxide supercritical fluid extraction, extraction temperature can be set at 30-60 ℃.
3, when carrying out carbon dioxide supercritical fluid extraction, extracting pressure can be set at 20-60MPa.
4, when carrying out carbon dioxide supercritical fluid extraction, the extraction time can be set at 1h or more than.
5, when carrying out carbon dioxide supercritical fluid extraction, entrainer can be ethanol more than 95% for ethanol or concentration, and consumption can be set at 0.1-1ml/g medicated powder.
6, when with Rotary Evaporators carbon dioxide supercritical fluid extraction liquid being carried out concentrating under reduced pressure recovery ethanol, temperature can be set at 45 ℃ or following.
7, when the medicinal residues behind the carbon dioxide supercritical fluid extraction are extracted, extracting solvent can be 30%-100% ethanol.
8, when the medicinal residues behind the carbon dioxide supercritical fluid extraction are extracted, extracting mode can be that heating and refluxing extraction, supersound extraction or percolation extract.
9, when ethanol extraction extractum is carried out the macroporous adsorbent resin enrichment, the post fat of use can be D-101 type or AB-8 type, and other are nonpolar, low pole or middle polarity macroporous adsorbent resin.
10, when ethanol extraction extractum is carried out the macroporous adsorbent resin enrichment, eluting solvent can be water or Different concentrations of alcohol, according to concentration of alcohol eluting successively from low to high.
By the extract that technique scheme prepares, have tangible tyrosinase activity inhibitory action and melanin and generate inhibitory action, can be for the preparation of pharmaceutical preparation, health food and the cosmetics of whitening and speckle dispelling.
When the pharmaceutical preparation that the said extracted thing is mixed into the Pear Power effect of dispelling spots and health food, except these extracts, in the scope of not overslaugh effect of the present invention, can suitably be mixed in other compositions of common pharmaceutical preparation and health food, for example as required: add adjuvant in crospolyvinylpyrrolidone, carboxymethylstach sodium, microcrystalline Cellulose, lactose, aspartame, the magnesium stearate etc., add adjuvant and coating adjuvant.Its type agent can be any pharmaceutically said dosage form, preferred pill, tablet, capsule or oral liquid.
When the cosmetics that the said extracted thing are mixed into the Pear Power effect of dispelling spots and external medicine preparation, except these extracts, in the scope of not overslaugh effect of the present invention, can suitably be mixed for as required in other compositions of skin preparations for extenal use such as common cosmetics, pharmaceuticals, for example: oil content, wetting agent, UV absorbent, antioxidant, surfactant, antiseptic, heat preserving agent, spice, water, alcohol, thickening agent etc.Extract of the present invention can be used for cosmetics and external medicine preparation, is particularly suitable for being widely used in the cosmetics.Its dosage form is so long as be used for the dosage form of skin and get final product, applicable to solution system, can dissolve any dosage form such as system, emulsification system, ointment, gel.In addition, it uses form also is arbitrarily, for example: astringent, emulsion, frost, facial film etc.
The pure natural plants extract that Chinese medicine compound extract of the present invention is dietotherapeutic, effect experiment subsequently will prove that Chinese medicine compound extract of the present invention has the tyrosinase activity inhibitory action and melanin generates the effect that suppresses.
The specific embodiment
Below in conjunction with specific embodiment, further set forth the present invention.These embodiment are interpreted as only being used for explanation the present invention and are not used in restriction protection scope of the present invention.After the content of having read the present invention's record, those skilled in the art can make various changes or modifications the present invention, and these equivalences change and modify and fall into claims of the present invention institute restricted portion equally.For convenience of explanation, abbreviate extract of the present invention as TQW below.
Embodiment 1 (experimentation that the TQW restraint of tyrosinase is active and melanin generates)
1 experiment material
Mice B16 melanoma cells (purchasing the Shanghai cell institute in the Chinese Academy of Sciences); Arbutin (purchasing the company in Sigma); DMEM culture medium (purchasing the company in Gibco); Hyclone (purchasing the company in Billab); Trypsin is purchased the company in Billab); Phosphate-buffered salt (purchasing China fir Golden Bridge company limited in Beijing); Two anti-(purchasing the company in Billab); DOPA (purchasing the company in Sigma); TritonX-100 (purchasing the company in Sigma); MTT (purchasing the company in Billab); DMSO (purchasing the company in Sigma); Microplate reader (thermoelectric Shanghai Instr Ltd.).
The preparation of 2 reagent
Getting arbutin faces the time spent and is diluted to the experiment desired concn with the DMEM culture medium.
Get TQW, use a small amount of anhydrous alcohol solution, be made into 1mg/ml with the dissolving of DMEM culture medium again, wherein concentration of alcohol is 0.8%, and regulating pH value is 7.0, filtration sterilization, and 4 ℃ of preservations, standby.
3 cell culture and exposed by the reagent thing
Get mice B16 melanoma cells, treat that cell grows to nearly fusion state, through 0.25% trypsinization, go down to posterity with the DMEM culture fluid that contains 10% calf serum, place CO
237 ℃ of incubators, 5%CO
2Cultivate in the saturated humidity environment.Same passage cell is taken from experiment each time.
3.1 experiment grouping
Blank group is to add cell culture fluid in the hole of inoculating cell not; Negative control group is to add cell culture fluid in the hole of having inoculated cell; The administration group is to add the culture fluid that contains variable concentrations TQW in the hole of having inoculated cell respectively; The arbutin group is to add the culture fluid that contains the variable concentrations arbutin in the hole of having inoculated cell respectively.
3.2 cell proliferation inhibition rate is measured
Selection grows to the cell of nearly fusion state, uses 0.25% trypsinization, adjusts cell concentration, with every hole 5 * 10
3Individual 96 orifice plates that are inoculated in, every hole amount of liquid 150 μ L, place 37 ℃, volume fraction is after cultivating 24h under the 5%CO2 condition, negative control group is changed liquid 1 time with fresh medium, administration group and arbutin group are subjected to the fresh medium of reagent thing to change liquid 1 time with adding, and arbutin, TQW concentration are established 10 μ g/ml, 50 μ g/ml, 100 μ g/ml respectively, and each concentration is established 12 multiple holes; Blank group only adds culture fluid and does not add cell; Each group all places 37 ℃, and volume fraction is 5%CO
2Cultivate 72h under the condition, 4h adds MTT 20 μ L/ holes before finishing, and hatches abandoning supernatant behind the 4h for 37 ℃, adds dimethyl sulfoxide (DMSO) 150 μ L/ holes, vibrates about 10min.Survey the OD value with microplate reader, wavelength is 490nm.
Cell proliferation inhibition rate %=[1-(administration group OD value-blank group OD value)/(negative control group OD value-blank group OD value)] * 100%
3.3 the tyrosinase activity suppression ratio is measured
Selection grows to the cell of nearly fusion state, uses 0.25% trypsinization, adjusts cell concentration, with every hole 5 * 10
3Individual 96 orifice plates that are inoculated in, every hole amount of liquid 150 μ L, place 37 ℃, volume fraction is after cultivating 24h under the 5%CO2 condition, negative control group is changed liquid 1 time with fresh medium, administration group and arbutin group are subjected to the fresh medium of reagent thing to change liquid 1 time with adding, and arbutin, TQW concentration are established 10 μ g/ml, 50 μ g/ml, 100 μ g/ml respectively, and each concentration is established 12 multiple holes; Blank group only adds culture fluid and does not add cell; Each group all places 37 ℃, and volume fraction is to cultivate 72h under the 5%CO2 condition, and the culture fluid that inclines is washed twice with PBS, add TritonX-10090 μ L/ hole, concussion 30min adds 0.2%L-dopa solution 60 μ L/ holes, hatch 20min for 37 ℃, survey the OD value with microplate reader, wavelength is 490nm.
Tyrosinase activity suppression ratio %=[1-(administration group OD value-blank group OD value)/(negative control group OD value-blank group OD value)] * 100%
3.4 the melanin content suppression ratio is measured
Select the exponential phase cell, use 0.25% trypsinization, adjust cell concentration and be inoculated in the culture bottle, place 37 ℃, volume fraction is 5%CO
2After cultivating 24h under the condition, negative control group is changed liquid 1 time with fresh medium, administration group and arbutin group are subjected to the fresh medium of reagent thing to change liquid 1 time with adding, and arbutin, TQW concentration are established 10 μ g/ml, 50 μ g/ml, 100 μ g/ml respectively, and each concentration is established 12 multiple holes; Blank group only adds culture fluid and does not add cell; Each group all places 37 ℃, and volume fraction is 5%CO
2Cultivate 72h under the condition, with 0.25% pancreatin harvesting, centrifugal (1500r/min, 5min) back abandoning supernatant adds the PBS buffer and regulates cell density to 10
5About/ml, each is drawn 1ml cell suspension and places 3 parallel tool plug pipes respectively, centrifugal (1500r/min, 5min) back abandoning supernatant, adding 200 μ l distilled water earlier suspends cell again, add 1ml ethanol then: ether solution (volume ratio is 1: 1) is to dissolve the opaque granule of non-melanocyte, at room temperature place 15min, centrifugal (3000r/min, 5min) and abandoning supernatant, adding 1ml mass fraction is 10% dimethyl sulfoxide, 1mol/LNaOH solution, places 80 ℃ of water-bath 30min, and cell mass is dissolved fully.Survey the OD value with microplate reader, wavelength is 490nm.
Melanin content suppression ratio %=[1-(administration group OD value-blank group OD value)/(negative control group OD value-blank group OD value)] * 100%
4 interpretations of result
4.1TQW the influence to the growth of B-16 cell
By table 1 as seen, the inhibitory action of TQW cell growth with compare with the concentration arbutin, the equal not statistically significant of difference (P＜0.05) illustrate that the cell growth inhibition of TQW is similar to traditional whitening agent arbutin, and cytotoxicity is lighter.
4.2TQW to B-16 cell tyrosinase activity and the synthetic influence of melanocyte
By table 2,3 as seen, three concentration of TQW all have the effect that significant restraint of tyrosinase is active and melanocyte is synthetic, and effect all is better than arbutin, illustrate that at restraint of tyrosinase active and melanocyte synthesizes aspect, TQW has more obvious effect than traditional whitening agent arbutin.
The mice B16 of table 1TQW melanoma cells inhibition of proliferation rate (
%)
With compare with the concentration arbutin,
△P＜0.05,
△ △P＜0.01
The suppression ratio of the mice B16 of table 2TQW melanoma cells tyrosinase activity (
%)
Compare with negative control,
*P＜0.05,
*P＜0.01; With compare with the concentration arbutin,
△P＜0.05,
△ △P＜0.01
The suppression ratio that the mice B16 of table 3TQW melanoma cells melanocyte synthesizes (
%)
Compare with negative control,
*P＜0.05,
*P＜0.01; With compare with the concentration arbutin,
△P＜0.05,
△ △P＜0.01
Embodiment 2 (the UVB of TQW induces the Pigmented inhibitory action experimentation of guinea pig skin)
1 experimental apparatus, reagent and laboratory animal
UVB ultraviolet Phototherapeutic instrument (Sigma company); 0.1%8-methoxypsoralen (8-MOP) tincture (Sigma company); VITAMIN C TABLET (Shijiazhuang Ouyi Pharmaceutical Co., Ltd); 90 of the healthy pure white Cavia porcelluss of regular grade, female, body weight 250-320g; Prepare the external used medicine that contains variable concentrations TQW with common substrate, contain the filling stomach medicine of variable concentrations TQW with the preparation of distilled water suspendible.
Pigmentation model copy and administration that 2UVB induces
Every guinea pig back is got 3 phase abscission zone territory (2 * 2cm
2), to wipe out the guinea pig back centre with curved scissors and become mildewed, electric shaver is shaved clean undercoat, uses the irradiation of SS-04B type ultraviolet phototherapy instrument, Burdick lamp configuration UVB light source, the spectrum peak of fluorescent tube is 310～315nm; Pre-irradiation is used the UVB ultraviolet radiation meter and is measured the irradiation light intensity, shines this zone 20min, every day irradiation dose 200mJ/cm
2, continuous 2 weeks, cumulative exposure 2800mJ/cm
2Each irradiation administration in back 2 hours, irradiation finishes the back and continues 2 weeks of administration, and the bark fetching skin tissue is done pathologic finding.
3 groupings
3.1 external experiment
Make blank with the normal guinea pig skin of back that does not shine ultraviolet for the 1st group; The 2nd group, modeling is coated with the employed substrate of preparation external used medicine outward, as own control; 3 and 4 groups, modeling is coated with the medicine that contains TQW outward, and concentration is respectively 0.5 and 1mg/ml; The 5th group, modeling is coated with 0.1%8-methoxypsoralen (8-MOP) tincture outward, as positive control.
3.2 experiment for oral administration
Make blank with the normal guinea pig skin of back that does not shine ultraviolet for the 1st group; The 2nd group, modeling is irritated stomach and is given distilled water 2ml, as own control; 3 and 4 groups, the medicine that stomach contains TQW is irritated in modeling, and dosage is 10 and the 20mg/kg body weight; The 5th group, modeling is irritated stomach and is given vitamin C, and dosage is the 20mg/kg body weight, as positive control.
4 colouring methods
4.1HE dyeing: (2cm * 2cm), routine paraffin wax section HE dyes from guinea pig back bark fetching skin specimen.
4.2Schmorl method dyeing
(1) section moves to washing.(2) with 0.037mol/L (1%) the iron chloride 30ml of fresh configuration, the mixed liquor of the 0.03mol/L of fresh configuration (1%) potassium ferricyanide 4mL and distilled water 6mL is handled 10min.(3) flowing water cleans.The result: melanocyte dyeing is yellowish-brown, black particle.
4.3Imokawa method dyeing
(1) (2cm * 2cm), with 0.1mol/LPBS liquid (pH6.8) flushing, the 1mol/L sodium bromide is hatched 5h for 37 ℃ from guinea pig back bark fetching skin specimen.(2) separate epidermis, corium.(3) epidermis fixedly 30min of the cold neutral formalin of 3.33mol/L (10%) is with 0.1mol/LPBS liquid (pH6.8) flushing 2 times.(4) with (pH6.8) 5h that dyes of 0.1% DOPA (0.1mol/LPBS liquid).The result: melanocyte is dyed pitchy.
5 om observations
5.1 contain the melanin granule cell counting: the dyeing of Schmorl method contains the melanin granule cell number with every square millimeter of stratum basale of the blind method every part of specimen of counting of net form eyepiece micrometer list under the high power lens.
5.2 DOPA positive cell counting: the dyeing of Imokawa method contains the dopa-positive melanocytes number with every square millimeter of stratum basale of the blind method every part of specimen of counting of net form eyepiece micrometer list under the high power lens.
6 interpretations of result
6.1 external experiment
By table 4 as seen, compare with the blank group, UVB model group guinea pig skin contains the melanin granule cell, DOPA positive cell number average obviously increases, and significant difference (P＜0.01) is arranged, the pigmentation skin model success that is similar to chloasma that this explanation UVB induces.TQW can obviously reduce guinea pig skin and contain melanin granule cell, DOPA positive cell number, and the effect of high concentration group is better than positive control drug 8-MOP.Illustrate that extract of the present invention has good whitening and speckle dispelling effect.
The influence that the guinea pig skin melanocyte of table 4 external TQW and melanocyte form
| Group | Contain melanin granule cell/mm 2 | The positive MC/mm of DOPA 2 |
| Blank group | 367.21±14.73 △△ | 169.43±11.02 △△ |
| The UVB model group | 476.01±21.40 ** | 240.12±12.58 ** |
| The TQW0.5mg/ml group | 419.09±15.26 **△△ | 201.67±8.43 **△△ |
| TQW 1mg/ml group | 370.68±16.20 △△ | 183.06±9.98 △△ |
| The 8-MOP group | 380.34±15.23 *△△ | 187.42±8.87 *△△ |
Compare with the blank group
*P＜0.05,
*P＜0.01; Compare with the UVB model group
△P＜0.05,
△ △P＜0.01
6.2 experiment for oral administration
By table 5 as seen, compare with the blank group, UVB model group guinea pig skin contains the melanin granule cell, DOPA positive cell number average obviously increases, and significant difference (P＜0.01) is arranged, the pigmentation skin model success that is similar to chloasma that this explanation UVB induces.TQW can obviously reduce guinea pig skin and contain melanin granule cell, DOPA positive cell number, and the effect of high concentration group is better than the positive control drug vitamin C.Illustrate that extract of the present invention has good whitening and speckle dispelling effect.
The influence that the table 5 guinea pig skin melanocyte of TQW for oral administration and melanocyte form
| Group | Contain melanin granule cell/mm 2 | The positive MC/mm of DOPA 2 |
| Blank group | 367.21±14.73 △△ | 169.43±11.02 △△ |
| The UVB model group | 476.01±21.40 ** | 240.12±12.58 ** |
| The TQW10mg/kg group | 429.3±14.35 **△△ | 215.76±8.41 **△△ |
| The TQW20mg/kg group | 375.74±12.27 △△ | 172.16±9.28 △△ |
| Vitamin C 20mg/kg group | 380.87±11.38 △△ | 176.09±10.41 △△ |
Compare with the blank group
*P＜0.05,
*P＜0.01; Compare with the UVB model group
△P＜0.05,
△ △P＜0.01
Embodiment 3 (preparation of tablet)
By weight, get TQW5～9 part, add 1～2 part of Pulvis Talci, 1～2 part of starch, 1～2 part of carboxymethyl starch sodium sprays into 95% ethanol moistening, wet granulation, drying adds 0.5～1% magnesium stearate, mix homogeneously, tabletting; Add 15% Opadry coating solution coating, 7% coating that increases weight gets Film coated tablets.
Embodiment 4 (preparation of capsule)
By weight, get TQW8～9 part, add 1～2 part of Pulvis Talci, spray into 95% ethanol moistening, wet granulation, drying adds 0.5～1% magnesium stearate, mix homogeneously, packing gets capsule.
Embodiment 5 (preparation of facial cream)
Preparation technology: (04) to (07) is mixed and heated to 80 ℃, adds (09), (10) to whole dissolvings, continuation adds (01), (02), (03) and (08) makes oil phase; (11) to (13) are added (14), be heated to 80 ℃ and make water, behind the biphase mixing and emulsifying, be cooled to adding (15) below 50 ℃, mixing gets final product.
Claims (2)
1. the Chinese medicine compound extract of Pear Power Eradicates speckle effect is characterized in that extracting method is as follows:
In weight portion, according to patrima villosa grass 4-6 part, Lignum Xanthoceratis kernel 4-6 part, Cortex Dictamni 2-4 part, Herba Agrimoniae 2-4 part, Radix Rhodiolae 2-4 part, Semen Cassiae 2-4 part, Semen Platycladi 2-4 part, Rhizoma Fagopyri Dibotryis 2-4 part, Herba Menthae 1-3 part, Radix Glycyrrhizae 1-2 part, it is an amount of to take by weighing each medical material, mix and pulverize, cross No. 2 sieves of pharmacopeia, medicated powder carries out carbon dioxide supercritical fluid extraction, be entrainer with ethanol, under certain technological parameter, extract pale brown color liquid, concentrating under reduced pressure reclaims ethanol, final pale brown color thickness grease; Again with the medicinal residues behind the previous step carbon dioxide supercritical fluid extraction, with extracting after the debita spissitudo soak with ethanol, extracting solution dry extractum, extractum utilizes macroporous adsorptive resins to carry out enrichment, makes brown ceramic powder under certain technological parameter; The pale brown color thickness grease that this powder and carbon dioxide supercritical fluid extraction are made mixes, and namely gets the Chinese medicine compound extract, wherein:
When the medicated powder of spending Herba Patriniae, Lignum Xanthoceratis kernel, Cortex Dictamni, Herba Agrimoniae, Radix Rhodiolae, Semen Cassiae, Semen Platycladi, Rhizoma Fagopyri Dibotryis, Herba Menthae, Radix Glycyrrhizae in vain is carried out carbon dioxide supercritical fluid extraction, extraction temperature is 30-60 ℃, extracting pressure is 20-60MPa, the extraction time be 1h or more than, entrainer dehydrated alcohol consumption is 0.1-1ml/g medicated powder, resolving pressure is 20MPa or following, and resolution temperature is 40 ℃ or following;
When the medicinal residues behind the carbon dioxide supercritical fluid extraction are extracted, extracting solvent is 30%-100% ethanol, consumption is 10 times of amounts for the first time, soaks 12 hours, and heating and refluxing extraction or supersound extraction or percolation extracted 2 hours, filter, in medicinal residues, add 5 times of amount 30%-100% ethanol again, extracted again 2 hours, filter, merging filtrate, drying under reduced pressure gets extractum;
When ethanol extraction extractum is carried out the macroporous adsorbent resin enrichment, extractum is added suitable quantity of water dilution back to be mixed with isopyknic D-101 type macroporous adsorbent resin and mixes thoroughly, be splined in the D-101 type macroporous adsorptive resins, applied sample amount is calculated as 1: 2 with the ratio of crude drug amount and amount of resin, static adsorption 30 minutes, carry out eluting with 20% ethanol of the water of 10 column volumes, 10 column volumes and 50% ethanol of 10 column volumes successively then, collect 50% ethanol elution, pulverize behind the drying under reduced pressure, make brown ceramic powder.
2. the described Chinese medicine compound extract of claim 1 is in the pharmaceutical preparation of preparation whitening and spot-eliminating or the application of health food.
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