CN105168359B - The application of radix fici simplicissimae and its extract - Google Patents
The application of radix fici simplicissimae and its extract Download PDFInfo
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- CN105168359B CN105168359B CN201510546810.1A CN201510546810A CN105168359B CN 105168359 B CN105168359 B CN 105168359B CN 201510546810 A CN201510546810 A CN 201510546810A CN 105168359 B CN105168359 B CN 105168359B
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- 239000000284 extract Substances 0.000 title claims abstract description 32
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 claims abstract description 24
- 230000002087 whitening effect Effects 0.000 claims abstract description 16
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 claims abstract description 10
- 229930003944 flavone Natural products 0.000 claims abstract description 10
- 150000002212 flavone derivatives Chemical class 0.000 claims abstract description 10
- 235000011949 flavones Nutrition 0.000 claims abstract description 10
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 101710147108 Tyrosinase inhibitor Proteins 0.000 claims abstract description 8
- 230000008021 deposition Effects 0.000 claims abstract description 8
- 239000002537 cosmetic Substances 0.000 claims abstract description 6
- 206010040829 Skin discolouration Diseases 0.000 claims abstract description 5
- 229940124200 Melanin inhibitor Drugs 0.000 claims abstract description 4
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- 235000019441 ethanol Nutrition 0.000 claims description 128
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- 239000000287 crude extract Substances 0.000 claims description 25
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- 239000003480 eluent Substances 0.000 claims description 13
- 238000002360 preparation method Methods 0.000 claims description 13
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- 230000002745 absorbent Effects 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 12
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- 229940126680 traditional chinese medicines Drugs 0.000 abstract description 3
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- BNGXYYYYKUGPPF-UHFFFAOYSA-M (3-methylphenyl)methyl-triphenylphosphanium;chloride Chemical compound [Cl-].CC1=CC=CC(C[P+](C=2C=CC=CC=2)(C=2C=CC=CC=2)C=2C=CC=CC=2)=C1 BNGXYYYYKUGPPF-UHFFFAOYSA-M 0.000 description 1
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- Cosmetics (AREA)
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Abstract
The present invention relates to technical field of traditional Chinese medicines, disclose radix fici simplicissimae and its extract and are preparing tyrosinase inhibitor, Melanin inhibitor, skin-lightening cosmetic and skin-whitening, preventing the application in skin melanin deposition.Radix fici simplicissimae and its extract have excellent tyrosinase inhibitory activity, especially radix fici simplicissimatis extractive has the more significant tyrosine inhibitory activity compared with existing tyrosinase inhibitor and whitening agent arbutin, having simultaneously prevents melanin deposition effect and higher free radical scavenging activity, containing more general flavone, can be preferably applied in beautifying whitening field.
Description
Technical field
The present invention relates to technical field of traditional Chinese medicines, particularly relate to the new opplication of radix fici simplicissimae and its extract.
Background technique
The colour of skin is that people examine one of standard of beauty.For Asia women, the pale colour of skin is even more that one kind is generally pursued.For this purpose,
Various products are come into being, such as: whitening products, antiultraviolet product and film product.Through studying, skin color is by Skin Cell
Contained in various pigments are comprehensive determines.In numerous pigments, melanin amount contained in melanocyte is most important decision
Factor.Further research confirms that tyrosinase (EC1.14.18.1) is the key enzyme of B16 cell.As a result, by inhibiting junket
Propylhomoserin enzymatic activity, to reduce melanin production, so that the skin effect for reaching whitening becomes the important evidence of research and development skin-protection product
One of (Chinese journals of practical medicine [J], 2005,21,1364-1366；Xiamen University's journal (natural science edition) [J], 2007,46,
274-282).Common whitening agent is such as in these products: quinhydrones, kojic acid, arbutin are several, these substances more or less have
There is comparable cytotoxicity.Therefore, the safety of tyrosinase inhibitor is to need to lay equal stress in whitening and skin-protecting R & D of complex
The factor of consideration, and the selection of Chinese herbal medicine is undoubtedly more easily esthetically acceptable to the consumers in this respect.
There are medicine in radix fici simplicissimae, alias Ficus hirta, cairo morningglory root or leaf, milk root, southern stilbene, multiple nationalitys such as Guangxi, Guangdong, Yunnan
With record, the ground such as main product Guangxi, Guangdong are known as " panacis quinquefolii radix ".Radix fici simplicissimae is Moraceae ficus species ficus simplicissima
(Ficushirtavahl.) root.Radix fici simplicissimae is used for health care civil, there is invigorating spleen to remove dampness, relieving cough and reducing sputum, clearing and activating the channels and collaterals
The effect of, in the civil usage history for having more than one thousand years in Guangdong.Zhang Baoxu of Peking University etc. (number of patent application:
200510002111.7) disclose new application of the radix fici simplicissimae in preparation hepatitis adjuvant therapy medicaments and health food.South
The Yuan Lixia (application number: 201010274140.X) of square medical university discloses a kind of pharmaceutical composition for preventing and treating vital myocarditis
Object and preparation method thereof.Wang Xiao equality (Wang Xiao equality, the experimental study of radix fici simplicissimae Aqueous extracts vitro antibacterial activity, Shi Zhenguo
Cure traditional Chinese medicines, 2010,21 (7): 1692-1693.) radix fici simplicissimae Aqueous extracts are reported to Escherichia coli, bacillus subtilis, golden yellow
Color staphylococcus all has inhibiting effect, and minimum inhibitory concentration is 1g/ml.Furthermore also have been reported that radix fici simplicissimatis extractive has
Anti-inflammatory, anti-acid and anti-aging effect.But mainly application is only limitted to above-mentioned several aspects to radix fici simplicissimae at present, mentions to it
Taking the other application of object to be worth also yet there are no in-depth study report.
Chinese patent CN103877283A discloses the Chinese medicine liquid of a kind of whitening spot-removing, toxin-expelling and face nourishing, to include the five fingers
10 kinds of Chinese medicinal composition compound decoctings including wild peach obtain, and can inhibit tyrosinase activity, and there is whitening spot-removing and toxin expelling to support
The effect of face.But the realization of the effect is established on the basis of entire Chinese medicine compound prescription collective effect, which does not disclose the five fingers
Wild peach or radix fici simplicissimatis extractive are individually able to suppress tyrosinase activity, and have the effect of skin whitening.
There is not the record of beautifying whitening related efficacy in Chinese medicine record in radix fici simplicissimae, has no five in the prior art yet
Refer to that wild peach is applied to the report of skin-lightening cosmetic as the purposes of tyrosinase inhibitor and radix fici simplicissimae.
Summary of the invention
In view of this, that the purpose of the present invention is to provide radix fici simplicissimaes and its extract is relevant in a series of beautifying whitenings
Using being mainly included in and prepare tyrosinase inhibitor, Melanin inhibitor, skin-lightening cosmetic and skin-whitening, anti-
The only new opplication in melanin deposition.
Record the field of Chinese medicines, radix fici simplicissimae the effect of be only limitted to strengthening the spleen and tonifying the lung, promoting the circulation of qi dampness removing, relaxing tendons and activating collaterals it
Function, the Chinese medicine of Lingnan area or the civil doctor of ethnic group be usually used in treat spleen deficiency edema, deficiency of food inability, pulmonary tuberculosis cough, night sweat,
The diseases such as leukorrhagia, postpartum agalorrhea, irregular menstruation, rheumatic arthralgia, oedema.And the present invention has found by creative research, radix fici simplicissimae
Extract has the ability preferably inhibited tyrosinase activity, can be used in the preparation of skin-whitening and related cosmetics.
Preferably, the radix fici simplicissimatis extractive is its water extract or alcohol extract.
It is highly preferred that radix fici simplicissimatis extractive of the present invention is prepared by the following:
Step 1, radix fici simplicissimae are added ethanol water after crushing and extract, and solvent acquisition radix fici simplicissimae is removed after extraction and is slightly mentioned
Object；
Step 2, radix fici simplicissimae crude extract add deionized water dissolving, then loading to macroporous absorbent resin, successively spend from
Sub- water, gradient concentration ethanol solution from low to high carry out gradient elution, collect 60v/v%-95v/v% concentration therein respectively
The eluent of ethanol elution, removed under reduced pressure second alcohol and water obtain radix fici simplicissimatis extractive.
Radix fici simplicissimatis extractive of the present invention can be each single concentration ethanol in 60v/v%-95v/v% concentration
The eluent of elution is also possible to their mixture.
In certain embodiments of the present invention, 60v/v%-95v/v% concentration ethanol therein is collected in step 2 respectively
The eluent of elution can be eluent (the alternatively referred to as 60v/v% alcohol elution, with lower class of 60v/v% ethanol elution
Together), the eluent of the eluent of 80v/v% ethanol elution, the eluent of 90v/v% ethanol elution, 95v/v% ethanol elution.
Preferably, being extracted as ultrasonic extraction described in step 1, stirring is extracted or circulated in countercurrent is extracted.
Preferably, the gradient concentration ethanol solution described in step 2 from low to high is the ladder of 20v/v%-95v/v% concentration
Spend concentration ethanol solution.It is highly preferred that the gradient concentration ethanol solution described in the step 2 from low to high is by 20v/v% second
Alcoholic solution, 30v/v% ethanol solution, 40v/v% ethanol solution, 60v/v% ethanol solution, 80v/v% ethanol solution, 90v/
The gradient concentration ethanol solution of v% ethanol solution and 95v/v% ethanol solution composition.In addition, in some specific realities of the invention
It applies in mode, selection that one of the gradient concentration ethanol solution from low to high can also be following:
(1) 20v/v% ethanol solution, 40v/v% ethanol solution, 60v/v% ethanol solution, 80v/v% ethanol solution,
95v/v% ethanol solution；
(2) 40v/v% ethanol solution, 60v/v% ethanol solution, 80v/v% ethanol solution；
(3) 30v/v% ethanol solution, 60v/v% ethanol solution, 90v/v% ethanol solution.
Preferably, the macroporous absorbent resin is AB-8 type macroporous absorbent resin, D301 type macroporous absorbent resin or X-5
Type macroporous absorbent resin.
Preferably, the extraction time of extraction described in step 1 is 0.5-2h, in certain specific embodiments of the invention
In, it can choose 0.5h, 1h or 2h.
Preferably, the mass volume ratio of radix fici simplicissimae and ethanol water is 1g:10-20mL in step 1, in the present invention
Some specific embodiments in, can be indicated with 40g and 400mL, 40g and 800mL, 20g and 400mL.
Preferably, the concentration of alcohol of ethanol water described in step 1 is 50v/v%-90v/v%, of the invention one
In a little specific embodiments, 50v/v%, 70v/v% or 90v/v% can choose.
Preferably, the general flavone content of the radix fici simplicissimatis extractive is 7.9-17.5%.
Preparation method of the present invention is obtained into radix fici simplicissimatis extractive and existing tyrosinase inhibitor and whitening agent bear
Fruit glycosides compares, and the tyrosinase inhibition rate of arbutin is 43%, and the tyrosinase of radix fici simplicissimatis extractive of the present invention
Inhibiting rate is in 45-69%, hence it is evident that higher than the inhibitory effect of existing product arbutin.
Meanwhile containing more general flavone in radix fici simplicissimatis extractive of the present invention, and removing free radical effect side
Face, radix fici simplicissimatis extractive of the present invention have the free radical scavenging activity of up to 65-77%, can be using in whitening product.
In addition, radix fici simplicissimatis extractive of the present invention is acted on zebrafish embryo, the results show that of the present invention five
Refer to that wild peach extract has inhibitory activity to zebra fish melanin production, skin melanin deposition can be prevented.Preferably, have
The radix fici simplicissimatis extractive of melanin production inhibitory activity is to obtain according to radix fici simplicissimatis extractive preparation method of the present invention
80v/v% concentration ethanol elutes position.
Radix fici simplicissimatis extractive of the present invention both can be applied to each application of the present invention separately as active material
In field, other active materials can also be cooperated to be used together.
From the above technical scheme, radix fici simplicissimae and its extract have excellent tyrosinase inhibitory activity, especially
It is that radix fici simplicissimatis extractive has the more significant tyrosine compared with existing tyrosinase inhibitor and whitening agent arbutin
Inhibitory activity, while having and prevent melanin deposition effect and higher free radical scavenging activity, it, can containing more general flavone
It is preferably applied in beautifying whitening field.
Detailed description of the invention
Fig. 1 show the microscopy figure of zebrafish embryo apparent black element inhibitory effect；Wherein 1-A indicates blank control microscopy
As a result；1-B is the microscopic examination result handled using radix fici simplicissimae 80v/v% alcohol elution.
Specific embodiment
The embodiment of the invention discloses radix fici simplicissimae and its new opplications of extract.Those skilled in the art can use for reference this
Literary content, is suitably modified realization of process parameters.In particular, it should be pointed out that all similar substitutions and modifications are to art technology
It is it will be apparent that they are considered as being included in the present invention for personnel.Application of the invention has passed through preferred embodiment
Be described, related personnel obviously can not depart from the content of present invention, in spirit and scope to product as described herein and side
Method is modified or appropriate changes and combinations, carrys out implementation and application the technology of the present invention.
For a further understanding of the present invention, below with reference to embodiment to radix fici simplicissimae provided by the invention and its extract
New opplication is described in detail.
Embodiment 1: the preparation of radix fici simplicissimatis extractive of the present invention
It by 40g radix fici simplicissimae crushed after being dried, is mixed with 400mL70v/v% ethanol solution, ultrasonic extraction 0.5 hour, point
Crude extract is separated out, after removed under reduced pressure solvent, obtains radix fici simplicissimae crude extract 2.6g.
Take radix fici simplicissimae crude extract 1.6g, after deionized water dissolving, be loaded to AB-8 resin packed column, successively spend from
Sub- water, 20v/v% ethyl alcohol, 40v/v% ethyl alcohol, 60v/v% ethyl alcohol, 80v/v% ethyl alcohol and 95v/v% ethanol gradient elution, point
Not Shou Ji 60v/v% ethyl alcohol, 80v/v% ethyl alcohol, 95v/v% ethyl alcohol eluent, removed under reduced pressure second alcohol and water, obtain the five fingers hair
Peach extract collects freezen protective at -20 DEG C.
Embodiment 2: the preparation of radix fici simplicissimatis extractive of the present invention
It by 20g radix fici simplicissimae crushed after being dried, is mixed with 400mL50v/v% ethanol solution, stirring is extracted 2 hours, separation
Crude extract out obtains radix fici simplicissimae crude extract 1.7g after removed under reduced pressure solvent.
Take radix fici simplicissimae crude extract 1.0g, after deionized water dissolving, be loaded to D301 resin packed column, successively spend from
Sub- water, 40v/v% ethyl alcohol, 60v/v% ethyl alcohol, 80v/v% ethanol gradient elution collect 60v/v% ethyl alcohol, 80v/v% respectively
The eluent of ethyl alcohol, removed under reduced pressure second alcohol and water obtain radix fici simplicissimatis extractive, collect freezen protective at -20 DEG C.
Embodiment 3: the preparation of radix fici simplicissimatis extractive of the present invention
By 40g radix fici simplicissimae crushed after being dried, being mixed with 800mL90v/v% ethanol solution, circulated in countercurrent is extracted 1 hour,
Crude extract is isolated, after removed under reduced pressure solvent, obtains radix fici simplicissimae crude extract 1.9g.
Take radix fici simplicissimae crude extract 1.5g, after deionized water dissolving, be loaded to X-5 resin packed column, successively spend from
Sub- water, 30v/v% ethyl alcohol, 60v/v% ethyl alcohol, 90v/v% ethanol gradient elution collect 60v/v% ethyl alcohol, 90v/v% respectively
The eluent of ethyl alcohol, removed under reduced pressure second alcohol and water obtain radix fici simplicissimatis extractive, collect freezen protective at -20 DEG C.
Embodiment 4: determination of total flavonoids in radix fici simplicissimatis extractive
Instrument: the general general T6 ultraviolet-visual spectrometer of analysis,
Measure sample: extract of the present invention is the radix fici simplicissimatis extractive of 60v/v% alcohol elution in embodiment 1；
The radix fici simplicissimatis extractive of 80v/v% alcohol elution, 95v/v% alcohol elution；
Reference extract be the radix fici simplicissimae crude extract collected according to 1 method of embodiment, wash position radix fici simplicissimae mention
The radix fici simplicissimae of object, the radix fici simplicissimatis extractive of 20v/v% alcohol elution and 40v/v% alcohol elution is taken to extract
Object；
Determination step: according to Pharmacopoeia of People's Republic of China (2010 editions) method, using rutin as standard items, the five fingers hair is measured
General flavone content in peach extract.Determination step are as follows: 1) standard curve making: precision measure reference substance solution 1ml, 2ml, 3ml,
4ml, 5ml, 6ml are set respectively in 25ml measuring bottle, and respectively plus 30% ethyl alcohol is to 6.0ml, add 5% sodium nitrite solution 1ml.It mixes, puts
Set 6 minutes, then plus 10% aluminum nitrate solution 1ml, shake up, place 6 minutes.Adding sodium hydroxide test solution 10ml, then plus 30% ethyl alcohol
It to scale, shakes up, places 15 minutes, using corresponding reagent as blank, with UV-VIS spectrophotometry, at the wavelength of 510nm
Absorbance is measured, using absorbance as ordinate, concentration is abscissa, draws standard curve.2) a certain amount of sample is taken to be placed in 25ml
It is other to measure absorbance in accordance with the law, simultaneously from " adding sodium nitrite solution 1ml " according to standard curve preparation step in volumetric flask
Test solution 3ml is taken, in addition to sodium hydroxide test solution is not added, remaining is ibid operated, and as blank, calculates radix fici simplicissimatis extractive
Middle general flavone content (in terms of rutin).The results are shown in Table 1 for determination of total flavonoids.
General flavone content in 1 radix fici simplicissimatis extractive of table
| Sample | General flavone content % |
| Radix fici simplicissimae crude extract | 5.1 |
| Water elution position radix fici simplicissimatis extractive | 0.3 |
| 20v/v% alcohol elution radix fici simplicissimatis extractive | 1.4 |
| 40v/v% alcohol elution radix fici simplicissimatis extractive | 3.6 |
| 60v/v% alcohol elution radix fici simplicissimatis extractive | 8.8 |
| 80v/v% alcohol elution radix fici simplicissimatis extractive | 17.5 |
| 95v/v% alcohol elution radix fici simplicissimatis extractive | 7.9 |
According to the above results as can be seen that preparing radix fici simplicissimatis extractive with high-content by the method for the invention
General flavone.
Embodiment 5: the detection of tyrosinase inhibitory activity
Test sample: according to the radix fici simplicissimae crude extract and difference of the preparation of embodiment 1, embodiment 2 and 3 method of embodiment
Concentration ethanol elutes the radix fici simplicissimatis extractive that position obtains, and has among these both included radix fici simplicissimatis extractive of the present invention,
Also include the radix fici simplicissimatis extractive of control.
Above-mentioned each sample is dissolved in deionized water, is configured to aqueous solution, for measuring tyrosinase inhibitory activity, with black bearberry
Glycosides is object of reference.Determination step are as follows: 70 μ L of soda acid buffer solution (pH6.8,30mM), 80 μ L of 1.66mM tyrosine solution, sample
80 μ L of solution is uniformly mixed, and is incubated 5 minutes or more in 37 DEG C of thermostatic water baths, and tyrosinase solution (125U/ml) 10 μ L is added,
After 37 DEG C of constant temperature 10min, with the absorbance A of microplate reader measurement 475nm wavelength475.Sample aqueous solution is replaced with deionized water,
Absorbance is equally measured as reference solution, the results are shown in Table 2.
Inhibitory activity against tyrosinase calculation formula are as follows: inhibiting rate (%)=(A0-(A475-B))/A0× 100%, wherein A0
It is the absorbance of reference solution, A475It is the absorbance of sample solution, B is the absorbance of sample blank solution.
The measurement result of 2 tyrosinase inhibitory activity of table
| Sample | Sample solution concentration mg/ml | Tyrosinase inhibition rate % |
| Embodiment 1 (radix fici simplicissimae crude extract) | 1.0 | 25 |
| Embodiment 1 (water elution position) | 1.0 | 15 |
| Embodiment 1 (20v/v% alcohol elution) | 1.0 | 11 |
| Embodiment 1 (40v/v% alcohol elution) | 1.0 | 22 |
| Embodiment 1 (60v/v% alcohol elution) | 1.0 | 57 |
| Embodiment 1 (80v/v% alcohol elution) | 0.5 | 57 |
| Embodiment 1 (95v/v% alcohol elution) | 0.5 | 45 |
| Embodiment 2 (30v/v% alcohol elution) | 1.0 | 27 |
| Embodiment 2 (60v/v% alcohol elution) | 1.0 | 45 |
| Embodiment 2 (90v/v% alcohol elution) | 1.0 | 62 |
| Embodiment 3 (40v/v% alcohol elution) | 1.0 | 15 |
| Embodiment 3 (60v/v% alcohol elution) | 1.0 | 56 |
| Embodiment 3 (80v/v% alcohol elution) | 1.0 | 69 |
| Arbutin | 1.0 | 43.0 |
As can be seen from Table 2, the crude extract obtained after radix fici simplicissimae is extracted has the inhibiting effect to tyrosinase activity, warp
After adsorbing resin separation, gained 60v/v% ethyl alcohol, 80v/v% ethyl alcohol, 90v/v% ethyl alcohol and 95v/v% alcohol elution are equal
It is able to achieve the inhibition to tyrosinase activity, and higher than the inhibiting effect of crude extract and arbutin.Wherein 80v/v% ethyl alcohol
It is most strong to the inhibiting effect of tyrosinase activity to elute position.
Embodiment 6: the detection of free-radical scavenging activity
Test sample: according to the radix fici simplicissimae crude extract and difference of the preparation of embodiment 1, embodiment 2 and 3 method of embodiment
Concentration ethanol elutes the radix fici simplicissimatis extractive that position obtains, and has among these both included radix fici simplicissimatis extractive of the present invention,
Also include the radix fici simplicissimatis extractive of control.
Its free-radical scavenging activity is measured with DPPH method, with VCFor object of reference.Determination step are as follows: accurately pipette extracting solution
2mL adds people 2mL DPPH ethanol solution (2 × 10 in 10mL tool plug test tube-4Mol/L), mix well, be stored at room temperature, 30min
The absorbance A of spectrophotometric determination 517nm wavelength is used afterwards517(using ethyl alcohol as reference)；Simultaneously measure 2mL DPPH solution with
The mixed absorbance A of 2mL ethyl alcoholcAnd 2mL extracting solution and the mixed absorbance A of 2mL ethanol solution0.It is measured in parallel three
It is secondary, it is averaged, calculates free radical scavenging activity according to the following formula.
Free radical scavenging activity=(1- (A517-A0)/Ac) × 100%.
The testing result of 3 DPPH method free-radical scavenging activity of table
Seen from table 3, the crude extract obtained after radix fici simplicissimae is extracted, is able to achieve the scavenging effect to DPPH free radical,
After absorption resin separation, gained 60v/v% ethyl alcohol, 80v/v% ethyl alcohol and 95v/v% alcohol elution are able to achieve pair
The removing of DPPH free radical, and improving a lot compared with the scavenging effect of crude extract, wherein 80v/v% ethanol elution
Position is most strong to the scavenging effect of free radical.
Embodiment 7: radix fici simplicissimatis extractive inhibits zebrafish embryo melanin to generate experiment
Radix fici simplicissimae 80v/v% alcohol elution prepared by embodiment 1, is dissolved in deionized water, is configured to 10mg/ml
Aqueous solution, for measuring the inhibiting effect to zebrafish embryo melanin production.Determination step is as follows: the raising of zebra fish with
The collection of embryo, Adult Zebrafish are raised in the circulating water culture system of laboratory, and 28.5 DEG C, light dark cycles are 14/10 small
When, raising is filtered through the circulatory system with water and is sufficiently aerated, and pH value keeps 8.0 or so, and daily feeding fairy shrimp three times, feeds weekly
Food 6 days.Select form normal, the larger and healthy sexal maturity zebra fish of individual is matched in hatching system by sex ration 1:2
It raises in 15L 10-15 bar glass aquarium.It turns off the light in oviposition evening before that day 22:00, morning next day, 8:00 system was automatic
It turns on light, starts to lay eggs under light stimulation, collect embryo.After the embryo of collection adequately to be cleaned to removal foul, in microscope
Under pick out the normal embryo of healthy development, using 96 orifice plates as test chamber, be separately added into radix fici simplicissimatis extractive in every hole
Aqueous solution is assigned randomly in 96 orifice plates by 1 embryo/hole.Radix fici simplicissimatis extractive aqueous solution to be measured is replaced daily and is stirred,
To ensure that active constituent is evenly distributed.Pigmentation assessment based on phenotype, observes the melanin inhibitory effect of zebrafish embryo,
Detect that the results are shown in Table 4, zebrafish embryo microscope map is as shown in Figure 1.
4 zebrafish embryo melanin inhibitory effect of table
As shown in Table 4,80% ethyl alcohol position of radix fici simplicissimatis extractive has the inhibition to zebrafish embryo melanin production
Effect, at the same it is high-visible in the microscopic examination result of Fig. 1, and there are a large amount of melanin depositions in blank control, and pass through the five fingers hair
The microscopic examination result of peach extract-treated is but without any melanin deposition.
The above description of the embodiment is only used to help understand the method for the present invention and its core ideas.It should be pointed out that pair
For those skilled in the art, without departing from the principle of the present invention, the present invention can also be carried out
Some improvements and modifications, these improvements and modifications also fall within the scope of protection of the claims of the present invention.
Claims (10)
1. radix fici simplicissimae alcohol extract prepare the application in skin-lightening cosmetic, the skin-lightening cosmetic being capable of skin whitening；It is described
Radix fici simplicissimae alcohol extract is prepared by following methods:
Step 1, radix fici simplicissimae are added ethanol water after crushing and extract, and solvent is removed after extraction and obtains radix fici simplicissimae crude extract；
Step 2, radix fici simplicissimae crude extract add deionized water dissolving, then loading to macroporous absorbent resin, successively with deionized water,
Gradient concentration ethanol solution from low to high carries out gradient elution, collects 60v/v%-95v/v% concentration ethanol therein respectively and washes
De- eluent, removed under reduced pressure second alcohol and water obtain radix fici simplicissimae alcohol extract.
2. applying according to claim 1, which is characterized in that be extracted as ultrasonic extraction described in step 1, stirring is extracted or adverse current
Circulation is extracted.
3. applying according to claim 1, which is characterized in that the gradient concentration ethanol solution described in step 2 from low to high is
The gradient concentration ethanol solution of 20v/v%-95v/v% concentration.
4. applying according to claim 3, which is characterized in that the gradient concentration ethyl alcohol described in the step 2 from low to high is molten
Liquid is by 20v/v% ethanol solution, 30v/v% ethanol solution, 40v/v% ethanol solution, 60v/v% ethanol solution, 80v/v% ethyl alcohol
The gradient concentration ethanol solution of solution, 90v/v% ethanol solution and 95v/v% ethanol solution composition.
5. applying according to claim 1, which is characterized in that the macroporous absorbent resin be AB-8 type macroporous absorbent resin,
D301 type macroporous absorbent resin or X-5 type macroporous absorbent resin.
6. applying according to claim 1, which is characterized in that the extraction time of extraction described in step 1 is 0.5-2h.
7. applying according to claim 1, which is characterized in that the quality volume of radix fici simplicissimae and ethanol water in step 1
Than for 1g:10-20mL.
8. applying according to claim 1, which is characterized in that the general flavone content of the radix fici simplicissimae alcohol extract is 7.9-
17.5%。
9. radix fici simplicissimae alcohol extract is preparing the application in tyrosinase inhibitor；The radix fici simplicissimae alcohol extract is by following preparation
Method prepares:
Step 1, radix fici simplicissimae are added ethanol water after crushing and extract, and solvent is removed after extraction and obtains radix fici simplicissimae crude extract；
Step 2, radix fici simplicissimae crude extract add deionized water dissolving, then loading to macroporous absorbent resin, successively with deionized water,
Gradient concentration ethanol solution from low to high carries out gradient elution, collects 60v/v%-95v/v% concentration ethanol therein respectively and washes
De- eluent, removed under reduced pressure second alcohol and water obtain radix fici simplicissimae alcohol extract.
10. radix fici simplicissimae alcohol extract is preparing the application in Melanin inhibitor, the Melanin inhibitor can
Prevent skin melanin deposition；The radix fici simplicissimae alcohol extract is prepared by following preparation method:
Step 1, radix fici simplicissimae are added ethanol water after crushing and extract, and solvent is removed after extraction and obtains radix fici simplicissimae crude extract；
Step 2, radix fici simplicissimae crude extract add deionized water dissolving, then loading to macroporous absorbent resin, successively with deionized water,
Gradient concentration ethanol solution from low to high carries out gradient elution, collects 60v/v%-95v/v% concentration ethanol therein respectively and washes
De- eluent, removed under reduced pressure second alcohol and water obtain radix fici simplicissimae alcohol extract.
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| 李南薇等.五指毛桃功能性成分抗氧化活性研究.《食品工业》.2013,第34卷(第6期),第127-130页. |
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