CN109091429A - A kind of Rosa roxburghii Tratt polyphenol extract and preparation method and the application in cosmetics - Google Patents
A kind of Rosa roxburghii Tratt polyphenol extract and preparation method and the application in cosmetics Download PDFInfo
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
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Abstract
The invention discloses a kind of Rosa roxburghii Tratt polyphenol extract and preparation method and the applications in cosmetics.Polyphenol content is not less than 33% in Rosa roxburghii Tratt polyphenol extract of the invention.Steps are as follows for preparation method: step 1: taking fruit of Grossularia burejensis Berger, drying and crushing after crossing 50 meshes, is added ethanol water and is heated to required temperature, extract extracting solution under ultrasonic power；Step 2: taking extracting solution, solvent is recovered under reduced pressure, concentrated by rotary evaporation obtains Rosa roxburghii Tratt polyphenol crude extract；Step 3: taking Rosa roxburghii Tratt polyphenol crude extract, fill column, the thick polyphenol solution of certain density Rosa roxburghii Tratt that will be made into ethyl alcohol with macroreticular resin, with column purification on sample solution, eluent is afforded with stripping liquid again, using concentration is dried under reduced pressure, vacuum drying obtains Rosa roxburghii Tratt polyphenol extract powder.The advantages that present invention has extraction process simple, strong operability, polyphenol is with high purity.The present invention also provides Rosa roxburghii Tratt polyphenol extract can be as the raw material of skin-lightening cosmetic as tyrosinase inhibitor.
Description
Technical field
The present invention relates to Rosa roxburghii Tratt polyphenol extracts and the preparation method and application thereof, answering especially in terms of skin-lightening cosmetic
With.
Background technique
Rosa roxburghii Tratt (Rosa roxburghii Tratt.) be rosaceae rose rosa roxburghii fruit also known as thatch pears,
Oblonga, send the spring to return at Wen Guangguo, is the distinctive wild plant resource in the Yunnan-Guizhou Plateau, there is the effect of digestion-promoting spleen-invigorating, astringing to arrest diarrhea.
The substances such as vitamin, polyphenol, flavones, organic acid, polysaccharide, amino acid and microelement rich in Rosa roxburghii Tratt, have very
High nutritive value.The study found that a variety of active ingredients in Rosa roxburghii Tratt, such as Rosa roxburghii Tratt polyphenol, Rosa roxburghii Tratt flavones, Polysaccharides from Rosa roxburghii all have
Anti-aging, effect that is anti-oxidant and removing free radical, prompt have potential application prospect in skin-lightening cosmetic development field.
However the research about Rosa roxburghii Tratt whitening field is in the starting stage at present, the extractive technique and efficiency of Rosa roxburghii Tratt polyphenol are still without system
Research, influence and white-skinned face function evaluation to tyrosinase activity have no research report, limit Rosa roxburghii Tratt as whitening additive
Follow-up study.The present invention is using national characters medicinal material Rosa roxburghii Tratt as raw material, and system is prepared for Rosa roxburghii Tratt polyphenol extract, and it is right to study its
The cell Proliferation of B16 and the influence of tyrosinase activity, to be Rosa roxburghii Tratt in skin-lightening cosmetic application theoretical foundation is provided.
With the raising of standard of living, cosmetics have become the necessity of people's daily life, also get over to the quality requirements of cosmetics
Come higher." advocating nature " becomes the Main Trends of The Development of countries in the world cosmetics, makes every effort to use natural plants in cosmetics
Raw material.
The patent of Publication No. CN102028780A provides a kind of ionization radiation protection new application of Rosa roxburghii Tratt flavones, passes through
The method of water extract-alcohol precipitation prepares Rosa roxburghii Tratt flavones, tests applied to cell and the ionization radiation protection of animal；Publication No.
The patent of CN107802543A provides a kind of Rosa roxburghii Tratt element and shines skin Biofibre facial mask and preparation method thereof, is homogenized using Single Roxburgh Rose Fruit
Ultrasonic extraction afterwards collects supernatant, and filter residue continues to ferment, and merges supernatant and fermentation liquid, carries out radicals scavenging experiment, super oxygen
Disproportionation enzymatic activity and bacteriostatic activity；Publication No. CN107028818A provides the method for preparation skin whitening Rosa roxburghii Tratt particle.It is public
The number of opening for the patent of CN105663496A provide a kind of scald containing aloe and Rosa roxburghii Tratt berry extract, burn drug composition and
It is applied, and squeezing is mixed with aloe extract after being impregnated using sucrose, but does not study ingredient and treatment scald in Fructus Rosae Normalis extract
Mechanism；The patent disclosure of Publication No. the CN 201710633848 a kind of formula and production method of Rosa roxburghii Tratt stoste facial mask, it is open
Number for CN 104523535A a kind of skin whitening, moisturizing cosmetics of patent disclosure and its method, but undeclared extract roxburgh rose root side
Method, effective component and whitening function mechanism；A kind of Rosa roxburghii Tratt of the patent disclosure of Publication No. CN106309629A, humulus grass, mulberry leaf
Mixed extract, but undetermined wherein active constituent content and whitening function mechanism.The patent of Publication No. CN101062002 is public
Skin care compositions of the cloth containing Fructus Rosae Normalis extract, there is a moisturizing, anti-aging effects；The current patent about Rosa roxburghii Tratt research, document
The whitening effective component and the mechanism of action of undeclared Fructus Rosae Normalis extract.Comprehensive existing patent and documents and materials, existing preparation
The Rosa roxburghii Tratt that technology obtains mentions polyphenol and takes object polyphenol content low, and application of the Rosa roxburghii Tratt polyphenol extract in skin-lightening cosmetic also has no phase
The research of pass is reported.
Summary of the invention
Regarding the issue above, the present invention provides a kind of Rosa roxburghii Tratt polyphenol extract and preparation method and in cosmetics
In application.The Rosa roxburghii Tratt polyphenol extract prepared by this method compared with prior art, has extraction process simple,
The advantages that strong operability, polyphenol is with high purity.
A kind of Rosa roxburghii Tratt polyphenol extract, wherein polyphenol content is not less than 33%.
A kind of preparation method of Rosa roxburghii Tratt polyphenol extract, comprising the following steps:
Step 1: taking fruit of Grossularia burejensis Berger, drying and crushing after crossing 50 meshes, is added needed for a certain amount of ethanol water is heated to
Temperature extracts extracting solution under certain ultrasonic power；
Step 2: taking extracting solution, solvent is recovered under reduced pressure, certain temperature concentrated by rotary evaporation obtains Rosa roxburghii Tratt polyphenol crude extract.
Step 3: Rosa roxburghii Tratt polyphenol crude extract is taken, fills column with macroreticular resin, the certain density Rosa roxburghii Tratt being made into ethyl alcohol is thick
Polyphenol solution, with column purification on certain sample solution concentration, sample solution flow rate and sample solution pH, then centainly to desorb liquid stream
Speed, stripping liquid concentration, stripping liquid pH afford eluent, and using concentration is dried under reduced pressure, vacuum drying obtains Rosa roxburghii Tratt polyphenol
Extract powder.
Preferably, the volume fraction of ethyl alcohol is 0%-100% in ethanol water in step 1.
Preferably, the volume fraction of ethyl alcohol is 80% in ethanol water in step 1.
Preferably, quality-volume ratio of fruit of Grossularia burejensis Berger and the ethanol water is 1:5-1:150 in step 1.
Preferably, quality-volume ratio of fruit of Grossularia burejensis Berger and the ethanol water is 1:100 in step 1.
Preferably, heating temperature described in step 1 is 25 DEG C -100 DEG C.
Preferably, heating temperature described in step 1 is 90 DEG C.
Preferably, ultrasonic power described in step 1 is 80-400W.
Preferably, ultrasonic power described in step 1 is 250W.
Preferably, the extraction time of extracting solution described in step 1 is 0.5h-4h.
Preferably, the extraction time of extracting solution described in step 1 is 80min, 40min.
Preferably, the extraction time of extracting solution described in step 1 is 1-4 times.
Preferably, the extraction time of extracting solution described in step 1 is 3 times.
The step 2 is implemented as follows: by the extracting solution of acquisition in 40-60 DEG C of rotary evaporation, removal is by step
Ethyl alcohol in rapid 1 obtained extracting solution obtains Rosa roxburghii Tratt polyphenol crude extract.
Preferably, the extracting solution in step 2 is in 40 DEG C of rotary evaporations, removal is passed through in the extracting solution that step 1 obtains
Ethyl alcohol obtains Rosa roxburghii Tratt polyphenol crude extract.
The step 3 is specially the Rosa roxburghii Tratt polyphenol crude extract for obtaining step 2, passes through macropore with certain sample solution concentration
Adsorb resin adsorption, successively with the water of certain pH certain flow rate and low-concentration ethanol be eluted to it is colourless after, successively with 40-95% it is dense
The ethanol gradient elution of degree collects the ethanol eluate of 40-95%.
Preferably, macroreticular resin described in step 3 be middle polarity or low pole macroreticular resin,
Preferably, macroreticular resin is one of ADS-17, HPD-600, AB-8 and NKA-9 or a variety of in step 3.
Preferably, the macroreticular resin in step 3 selects AB-8 resin.
Preferably, the loading flow velocity in step 3 is 1-5mL/min.
Preferably, the loading flow velocity in step 3 is 3mL/min.
Preferably, the sample solution concentration in step 3 is 0.6-1.0mg/mL.
Preferably, the sample solution concentration in step 3 is 0.72mg/mL.
Preferably, the sample solution pH value in step 3 is 4-8.
Preferably, the sample solution pH value in step 3 is 5.59.
Preferably, the desorption flow velocity in step 3 is 1-5mL/min.
Preferably, the desorption flow velocity in step 3 is 2.69mL/min.
Preferably, the stripping liquid concentration in step 3 is 50%-90%.
Preferably, the stripping liquid concentration in step 3 is 69.4%.
Preferably, the stripping liquid pH value in step 3 is 4-8.
Preferably, the stripping liquid pH value in step 3 is 6.3.
The step 3 is dried under reduced pressure step specifically: by the extracting solution of acquisition in 40-60 DEG C of rotary evaporation, removal
Ethyl alcohol in de- liquid, then after vacuum dried or frozen drying, obtain Rosa roxburghii Tratt polyphenol extract.
Preferably, the drying means of Rosa roxburghii Tratt polyphenol extract is vacuum drying.
Rosa roxburghii Tratt polyphenol extract is prepared, after detecting its purity, the Rosa roxburghii Tratt polyphenol extract for detecting various concentration is black to mouse
The influence of melanoma B16 cell, mtt assay detect Rosa roxburghii Tratt polyphenol extract to the shadow of Murine melanoma B16 cells proliferation activity
It rings, inverted microscope observes its influence to cellular morphology and quantity, and dopa oxidase method detects its shadow to tyrosinase activity
It rings.The Rosa roxburghii Tratt polyphenol extract of various concentration has certain inhibiting effect to B16 cell Proliferation and tyrosinase activity, and is in
Now significant concentration dependent (P < 0.5), high dose Rosa roxburghii Tratt polyphenol extract can obviously reduce the quantity of B16 cell, increase thin
Born of the same parents' fragment；After drug-treated 48h, Rosa roxburghii Tratt polyphenol extract is to the IC for inhibiting Murine melanoma B16 cells proliferation50Respectively
134.16 μ g/mL, to the IC inhibited tyrosinase activity50Respectively 82.42 μ g/mL, and Rosa roxburghii Tratt polyphenol extract is black to mouse
The inhibiting effect of melanoma B16 cell tyrosinase is better than positive control VC.
The present invention also provides Rosa roxburghii Tratt polyphenol extract to Murine melanoma B16 cells be proliferated and tyrosinase activity it is equal
There is certain inhibiting effect, therefore the present invention also provides Rosa roxburghii Tratt polyphenol extracts to make up as tyrosinase inhibitor in whitening
Application in product.Experimental data provided by the invention shows that Rosa roxburghii Tratt polyphenol extract provided by the invention has tyrosinase
Good inhibiting effect, the IC inhibited tyrosinase activity when acting on 48h50Value is 134.16 μ g/mL, is significantly better than positive control
Product VC.Rosa roxburghii Tratt polyphenol extract has potential development prospect as tyrosinase inhibitor.
The advantages of the present invention are as follows:
(1) present invention heats extraction by ethyl alcohol and macroporous resin purification method obtains Rosa roxburghii Tratt using fruit of Grossularia burejensis Berger as raw material
Polyphenol extract.The preparation method rational technology of Rosa roxburghii Tratt polyphenol extract of the present invention, simple and easy to do, production cost is low,
The yield and purity is high of polyphenol compound, can be used for large batch of industrialized production, have a vast market application prospect.
(2) Rosa roxburghii Tratt polyphenol extract produced by the present invention has the tyrosinase activity of Murine melanoma B16 cells
Good inhibiting effect, tyrosinase inhibitory activity are better than positive reference substance VC, Rosa roxburghii Tratt polyphenol extract provided by the invention
It can be used for preparing skin-lightening cosmetic.
Detailed description of the invention
Fig. 1 (a): four kinds of macroporous absorbent resins are to the adsorption curve of the thick polyphenol extract of Rosa roxburghii Tratt, the wherein class of macroreticular resin
Type includes AB-8, HPD-600, ADS-17 and NKA-9.
Fig. 1 (b): four kinds of macroporous absorbent resins are to the desorption curve of the thick polyphenol extract of Rosa roxburghii Tratt, the wherein class of macroreticular resin
Type includes AB-8, HPD-600, ADS-17 and NKA-9.
Influence of Fig. 2 (a) loading flow velocity to the thick polyphenol adsorption effect of Rosa roxburghii Tratt, wherein loading flow velocity 1-5mL/min.
Influence of Fig. 2 (b) sample solution concentration to the thick polyphenol adsorption effect of Rosa roxburghii Tratt, wherein sample solution concentration is 0.6-1.0mg/
mL。
Influence of Fig. 2 (c) sample solution pH to the thick polyphenol adsorption effect of Rosa roxburghii Tratt, wherein sample solution pH is 4-8.
Fig. 3 (a) desorbs influence of the flow velocity to the thick polyphenol desorption effect of Rosa roxburghii Tratt.
Influence of Fig. 3 (b) stripping liquid concentration to the thick polyphenol desorption effect of Rosa roxburghii Tratt.
Influence of Fig. 3 (c) stripping liquid pH to the thick polyphenol desorption effect of Rosa roxburghii Tratt.
The influence of the murine melanoma cells B16 survival rate of Fig. 4 Rosa roxburghii Tratt polyphenol.
Influence (× 10) of Fig. 5 Rosa roxburghii Tratt polyphenol to mouse B16 cellular morphology.
Fig. 6 Rosa roxburghii Tratt polyphenol is on the active influence of mouse B16 cell tyrosinase.
Specific embodiment
The preparation of 1 Rosa roxburghii Tratt polyphenol crude extract of embodiment:
Experimental material: ethyl alcohol；Natrium carbonicum calcinatum；Forint phenol reagent；Gallic acid standard items；Macroporous absorbent resin ADS-
17、HPD-600、AB-8、NKA-9。
Electronic balance；UV-1800 type ultraviolet-uisible spectrophotometer；Thermostat water bath；Supersonic cleaning machine；Chromatographic column.
Thick polyphenol in fruit of Grossularia burejensis Berger is extracted using ultrasonic ethanol immersion:
Dry fruit of Grossularia burejensis Berger is crushed, 50 meshes are crossed.Rosa roxburghii Tratt powder is weighed, 70% ethyl alcohol is added in liquid-to-solid ratio 100:1,
Ultrasonic power 250W, 30 DEG C of ultrasonic extraction 40min.Filtering, concentration, obtains thick polyphenol medicinal extract.Using gallic acid as standard items, use
Folin-Ciocaltean method measures Rosa roxburghii Tratt Determination of Polyphenols.
The Quality Detection of Rosa roxburghii Tratt polyphenol crude extract:
The purity of 1 Rosa roxburghii Tratt polyphenol crude extract of table
Through detecting, the purity that Rosa roxburghii Tratt dry fruit extracts polyphenol is 7.40%, polyphenol purity in Rosa roxburghii Tratt polyphenol crude extract
It is not high, it is subsequent that Rosa roxburghii Tratt polyphenol extract is purified using macroreticular resin absorbing method.
The purifying of 2 Rosa roxburghii Tratt polyphenol crude extract of embodiment
For the purity and content for improving polyphenol in Fructus Rosae Normalis extract, Fructus Rosae Normalis extract is carried out using macroreticular resin absorbing method
Purifying.
The absorption property of different macroreticular resins is investigated: resin being filtered dry, precision weighs 1.0g in 250mL stuffed conical flask
In.The thick polyphenol of 1g is taken to be dissolved in 200mL distilled water, measuring its polyphenol concentration is C0, it is added in the conical flask of macroreticular resin
Each 50mL.30 DEG C of water-baths for 24 hours, are periodically rocked in thermostat water bath.Period Aspirate supernatant at regular intervals measures extinction
Degree, obtaining remaining polyphenol concentration in solution is C1.Each macroporous absorbent resin is calculated according to formula (1) to inhale the static state of Rosa roxburghii Tratt polyphenol
Attached amount:
Q in formulasIt indicates adsorbance (mg/g)；C0Indicate polyphenol concentration (mg/mL) in thick polyphenol solution；C1It indicates after drawing
Remaining polyphenol concentration (mg/mL) in solution；V indicates crude extract volume (mL)；M indicates resin quality (g).
As shown in Figure 1,4 kinds of macroporous absorbent resins all have biggish desorption rate at desorption initial stage, and basic in 2h
Reach desorption balance.Wherein, the desorption efficiency of AB-8 resin is maximum, can reach 91.47%, ADS-17 and takes second place, desorption efficiency is
The desorption efficiency of 87.55%, NKA-9 and two kinds of polar resins of HPD-600 are lower, are 71.78% and 63.72%.Comprehensively consider 4 kinds
The static adsorbance and desorption efficiency of macroporous absorbent resin, it can be found that AB-8 resin can not only adsorb in a short time it is biggish more
Phenol amount, and desorption efficiency is high, desorption rate is fast.So the adsorbent material that this experiment uses AB-8 resin to purify as Rosa roxburghii Tratt polyphenol,
Further studied.
Rosa roxburghii Tratt polyphenol crude extract purifying process single factor exploration: thick polyphenol medicinal extract is dissolved with ethyl alcohol, AB-8 macroreticular resin
Column loading is filled, stationary phase answers factor, successively investigates sample solution concentration (0.6,0.7,0.8,0.9,1.0mg/mL), sample solution (pH
4,5,6,7,8), loading flow velocity (1,2,3,4,5mL/min), eluate concentration (50,60,70,80,90%), eluent pH
(4,5,6,7,8), eluent flow rate (1,2,3,4,5mL/min) are eluted, obtain eluent, wave using Rotary Evaporators
Solid carbon dioxide point, vacuum drying obtain Rosa roxburghii Tratt polyphenol powder, measure Rosa roxburghii Tratt polyphenol purity using Folin-Ciocaltean method.
By Fig. 2 (a) it is found that adsorbance is maximum when loading flow velocity is 1mL/min, reach 8.17mg/g.When loading liquid stream
When speed is gradually increased, the adsorbance of Rosa roxburghii Tratt polyphenol reduces therewith, and when flow velocity is greater than 3mL/min, adsorbance reduces more aobvious
It writes.When illustrating that loading flow velocity is larger, it will cause Rosa roxburghii Tratt polyphenol and not yet outflowed by resin adsorption, cause adsorbance smaller；
And loading flow velocity it is too small when, then it is too long to will lead to the operating time, increase cost, and when flow velocity is 3mL/min, adsorbance according to
So 7 or more.Therefore choosing 3mL/min is the horizontal flow velocity of response surface 0.
By Fig. 2 (b) it is found that when sample solution concentration be lower than 0.8mg/mL when, adsorbance with the increase of sample solution concentration and
It significantly increases, when sample solution concentration is greater than 0.8mg/mL, adsorbance tends to be steady.Reach when sample solution concentration is 1mg/mL
Maximum value 8.16mg/g, there is downward trend later.Illustrate that, when sample solution concentration is too low, polyphenol does not connect sufficiently with resin in solution
Touching is sucked out, to cause the phenomenon of leakage；When sample solution concentration is excessively high, impurity increases in solution, generates competition with polyphenol and closes
System, causes adsorbance smaller, while excessive sample solution concentration is likely to result in the generation of precipitating.Comprehensively consider, chooses
0.8mg/mL is the horizontal sample solution concentration of response surface 0.
By Fig. 2 (c) it is found that when sample solution concentration increases with the raising of pH, reach maximum value when pH is 6
8.17mg/g, later as reduced trend is presented in the raising of pH.Polyphenol is mainly based on phenolic acid class, so Rosa roxburghii Tratt polyphenol solution
In certain acidity, and the height of pH can cause the variation of polyphenol existence form.It can be seen that being conducive in acid condition big
Absorption of the macroporous adsorbent resin to Rosa roxburghii Tratt polyphenol.PH is 5.9 to sample solution concentration after measured, close with the pH value of maximum adsorption effect,
Therefore the horizontal pH of response surface 0 selects the sample solution not made adjustments.
By Fig. 3 (a) it is found that with the increase for desorbing flow velocity, corresponding desorption efficiency is gradually decreased, and desorption flow velocity is
Desorption efficiency when 1mL/min is 92.14%, and the desorption efficiency when desorbing flow velocity and being 5mL/min is only 80.15%.This may
It is because stripping liquid and macroporous absorbent resin time of contact are shorter, cause polyphenol can not be from macroporous absorption when desorption flow velocity is larger
It is so abundant that desorb in resin, ultimately cause the reduction of desorption efficiency.But lower flow velocity will cause time cost substantially
Rise, desorption efficiency can still reach 85% or more when 3mL/min as seen from the figure.So selecting 3mL/min is the horizontal solution of response surface 0
Imbibition flow velocity.
By Fig. 3 (b) it is found that desorption efficiency increases with the increase of stripping liquid concentration when stripping liquid concentration is lower than 70%,
Reach maximum desorption efficiency 92.51% in concentration for 70%.Then reduce with desorption efficiency with the increase of desorption concentration, works as solution
Inhaling concentration for 90% is that desorption efficiency is only 79.14%.Therefore the concentration of the horizontal stripping liquid of response surface 0 is adjusted to 70%.
By Fig. 3 (c) it is found that when stripping liquid pH is lower than 6, with the increase of stripping liquid pH, corresponding desorption efficiency is also therewith
Increase, when pH=6 reaches maximum desorption efficiency 93.12%.When pH is greater than 6, desorption efficiency is reduced with the increase of pH.This may
It is related that acidity is presented with Rosa roxburghii Tratt polyphenol.Therefore selecting pH=6 is that response surface 0 is horizontal.
Response phase method optimizes macroreticular resin to the adsorption and desorption of the thick polyphenol of Rosa roxburghii Tratt: according to center combination (Box-
Benhnken) experimental design principle, using statistical software Design-Expert, with A1(loading flow velocity, mL/min), B1(on
Sample liquid concentration, mg/mL), C1(sample solution pH) three factors are the independent variable in absorbing process, are worth in response with adsorbance；
With A2(desorption flow velocity, mL/min), B2(stripping liquid concentration, %), C2(stripping liquid pH) three factors are oneself in desorption technique
Variable is worth in response with desorption efficiency.Specific such as table 2.
2. adsorption and desorption technique independent variable factor of table coding and level
3 AB-8 resin Dynamic Adsorption of table and desorption Optimum Experiment result
4 response surface curved surface variance analysis of table
Note: 0.05 remarkable effect of * P <；The effect of 0.01 highly significant of * P <；The extremely significant effect of * * P < 0.001.
Note:*P < 0.05significant effect；* P < 0.01very significant effect；* * PP <
0.001extremely significant effect.
If table 3 adsorbs shown in Optimum Experiment result items, with the increase of each factor, adsorbance increases simultaneously.With loading
Liquid pH (C1), sample solution concentration (B1), loading flow velocity (A1) increase, there is maximum in adsorbance, hereafter, with it is each because
There is different degrees of decline in the increase of element, adsorbance.Forecast analysis is carried out by regression model, optimal Dynamic Adsorption condition is upper
Sample liquid flow velocity 2.65mL/min, sample solution concentration 0.72mg/mL, sample solution pH=5.59, adsorbance predicted value are 8.22mg/g.
If table 3 desorbs shown in Optimum Experiment result items, with the increase of each factor, desorption efficiency increases simultaneously.With desorption
Flow velocity (A2), stripping liquid pH (C2), stripping liquid concentration (B2) increase, there is maximum in desorption efficiency, hereafter, with it is each because
There is different degrees of decline in the increase of element, desorption efficiency.Forecast analysis is carried out by regression model, optimal dynamic desorption condition is solution
Imbibition flow velocity 2.69mL/min, stripping liquid concentration 69.4%, stripping liquid pH=6.3, desorption efficiency predicted value are 93.26%.
Each experimental group P value as shown in 4 response surface curved surface variance analysis of table, shows that response surface analysis experiment result of the present invention can
Letter.
Process certification research: the purification condition of face optimization according to response, this experiment improved conditions, adsorption conditions are loading
Flow velocity 2.6mL/min, sample solution concentration 0.7mg/mL, sample solution pH are initial pH=5.9；Desorption condition is desorption flow velocity
2.7mL/min, stripping liquid concentration 69%, stripping liquid pH=6.3 carry out verification test, 3 groups of samples of parallel loading, average adsorption
Amount is 8.11mg/g, and deviation 1.3%, average desorption efficiency is 94.39%, deviation 1.2%, illustrates to adsorb binomial fitting effect
Fruit is good, with a high credibility.
According to the process conditions after optimization, purification experiment is carried out to Rosa roxburghii Tratt polyphenol, experimental result is as shown in table 5, and Rosa roxburghii Tratt is more
The purity of phenol increases 36.33% by original 7.40%, is 4.9 times before purification.The rate of recovery of Rosa roxburghii Tratt polyphenol is
20.37%.
Influence of 5. purifying process of table to Rosa roxburghii Tratt polyphenol purity and the rate of recovery
| Predicted value | Measured value | RSD value | |
| Adsorbance(mg/g) | 8.22 | 8.11±0.19 | 1.3% |
| Desorption efficiency(%) | 93.26 | 94.39±2.21 | 1.2% |
| Before absorption | After desorption | The rate of recovery | |
| Polyphenol purity(%) | 7.40±0.39 | 36.33±2.43 | 20.37% |
The present invention simulate by Design-Expert software progress Box-Behnken center combination experimental design excellent
Change, is purified according to the technique after optimization, the purity of polyphenol is increased by original 7.40% in Rosa roxburghii Tratt polyphenol extract
36.33%, for 4.9 times before purification.
Influence of the 3 Rosa roxburghii Tratt polyphenol extract of embodiment to murine melanoma cells B16 survival rate
The present invention is proliferated Murine melanoma B16 cells using thiazolyl blue (MTT) method detection Rosa roxburghii Tratt polyphenol extract
It influences, it is therefore intended that find out a suitable safe-dosaging limits.
Laboratory apparatus: BLX-800 microplate reader (U.S. BIO-TEK)；Carbon dioxide incubator (U.S. Thermo)；Centrifuge
(Hunan Xiang Yi laboratory apparatus development corporation, Ltd.)；Inverted microscope (Japanese Nikon)；Biohazard Safety Equipment (U.S. Thermo)；
Electric-heated thermostatic water bath (Shanghai Ke Xi test apparatus factory)；High-pressure sterilizing pot (Japanese TOMY)
Experimental material: RPMI-1640 culture medium；Fetal calf serum；Pancreatin；PBS；Dimethyl sulfoxide；Methyl thiazoly tetrazolium assay；
L-DOPA；TritonX-100
The present invention is proliferated Murine melanoma B16 cells using thiazolyl blue (MTT) method detection Rosa roxburghii Tratt polyphenol extract
It influences.
Rosa roxburghii Tratt polyphenol extract provided by the invention is frozen with the mother liquor that culture medium is configured to 1000 μ g/mL spare.It takes
The B16 cell of logarithmic growth phase is adjusted to cell density to 3.5 × 10 with culture medium4A/mL, by cell inoculation in 96 orifice plates
In, 100 μ L of every hole, 37 DEG C, 5%CO2It is incubated for for 24 hours under saturated humidity environment.It is dense that different ends are added into the culture hole of detection group
The Rosa roxburghii Tratt polyphenol extract provided by the invention of degree, so that the final concentration of Rosa roxburghii Tratt polyphenol extract in every hole is respectively as follows: 40,80,
160,320 μ g/mL, and set up control group: media alone group, positive drug control group (VC, concentration: 90 μ g/mL) and blank pair
According to group (culture medium), it is placed in incubator and acts on 48h.Add 20 μ L concentration to be 5mg/mL MTT solution to every hole, is acted in incubator
Carefully absorbs the supernatant in culture hole after 4h, then 100 μ L DMSO are added to every hole, shakes 10min, is completely dissolved crystallization
Afterwards, it is placed in microplate reader, selection Detection wavelength is 492nm, measures absorbance value, and calculate cell survival rate, cell survival
The calculation formula of rate is as follows:
Test results are shown in figure 4, thin using the Rosa roxburghii Tratt polyphenol extract processing mouse melanoma B16 of various concentration
Born of the same parents, mtt assay detect the proliferation activity of cell.As shown in figure 4, Rosa roxburghii Tratt polyphenol extract provided by the invention is to B16 cell Proliferation
Inhibiting effect is all had, and is in certain concentration dependent (P < 0.5), is 40 μ g/mL, 80 μ g/mL, 160 μ g/mL in concentration,
The survival rate for making B16 melanoma cells when 320 μ g/mL is respectively 84.27%, 80.84%, 45.34%, 10.37%.With
GrapHpad Prism 5.0, which calculates Rosa roxburghii Tratt polyphenol extract, leads to the inhibition concentration (IC of cell survival rate decline 50%50) be
134.16μg/mL.Wherein the vitamin C (VC) of 90 μ g/mL is used as positive control, makes the survival of Murine melanoma B16 cells
Rate is 54.29%, illustrates that Rosa roxburghii Tratt polyphenol extract of the invention can be added in cosmetics and uses.
Influence of the 4 Rosa roxburghii Tratt polyphenol extract of embodiment to murine melanoma cells B16 tyrosinase activity
Tyrosinase is the major rate-limiting enzyme of melanin genesis, by the activity for inhibiting tyrosinase, it is possible to reduce melanocyte
Synthesis.The evaluation method of white-skinned face function evaluation has: biochemical process, cell biology method and normal individual trial test.The present invention adopts
With the white-skinned face function of cell biology method evaluation Rosa roxburghii Tratt polyphenol extract.The present invention measures mouse melanin using dopa oxidase method
Tumor B16 cell tyrosine enzymatic activity.
The cell of logarithmic growth phase is adjusted to cell density to 3.5 × 10 with culture medium4A/mL, by cell inoculation in
In 96 orifice plates, 100 μ L of every hole, 37 DEG C, 5%CO2It is incubated for for 24 hours under saturated humidity environment.It is added not into the culture hole of detection group
With the Rosa roxburghii Tratt polyphenol extract provided by the invention of final concentration, make in every hole Rosa roxburghii Tratt polyphenol extract concentration be respectively 40,80,
160,320 μ g/mL, and set up control group: media alone group, positive drug control group (VC, concentration: 90 μ g/mL) and blank pair
According to group (culture medium), it is placed in incubator and acts on 48h.Medical fluid is sucked, after washing 2 times with PBS, every hole adds 1%TritonX-100 solution
100 μ L are put into rapidly -80 DEG C of refrigerators and freeze 1h, and being subsequently moved to melt at room temperature makes cell be completely severed, decompose, pre- at 37 DEG C
100 μ L of 0.1%L-DOPA is added in every hole after temperature, surveys A under 37 DEG C of water-bath 2h, postposition microplate reader492, absorbance value is measured, and
Cell survival rate is calculated, in triplicate, the calculation formula of cell survival rate is as follows for experiment:
Test results are shown in figure 5, and Rosa roxburghii Tratt polyphenol extract provided by the invention is to Murine melanoma B16 cells junket ammonia
Phytase activity all has inhibiting effect, and is in certain concentration dependent (P < 0.5).Rosa roxburghii Tratt polyphenol provided by the invention extracts
After object handles Murine melanoma B16 cells 48h, make when concentration is 40 μ g/mL, 80 μ g/mL, 160 μ g/mL, 320 μ g/mL
B16 melanoma cell tyrosinase activity is reduced to 95.96%, 46.50%, 9.78%, 2.91% respectively, is computed, Rosa roxburghii Tratt
After polyphenol handles 48h, IC50Keep mouse black wherein the vitamin C (VC) of 90 μ g/mL is used as positive control for 82.42 μ g/mL
Melanoma B16 cell tyrosine enzymatic activity is reduced to 72.20%, and Rosa roxburghii Tratt polyphenol extract is better than the inhibitory effect of tyrosinase
VC, the exploitation for prompting Rosa roxburghii Tratt polyphenol extract to can be used as cosmetic additive agent for subsequent skin-lightening cosmetic.
After inverted microscope observes the processing of Rosa roxburghii Tratt polyphenol extract, Murine melanoma B16 cells form and density change
Become situation, as a result as shown in Figure 6.Normal group cell growth state is good, and form is normal, and adherent secured, degrees of fusion is good.It is different dense
After the Rosa roxburghii Tratt polyphenol extract processing of degree, cell density is reduced, and different degrees of variation, degrees of fusion occurs in cellular morphology
It reduces, cannot mutually be fused into reticular structure, with the increase of drug dose, the quantity of cell is significantly reduced, and cell fragment increases
It is more.The result is consistent with the result of MTT, further demonstrates increasing of the Rosa roxburghii Tratt polyphenol extract to Murine melanoma B16 cells
Grow inhibiting effect.
Claims (10)
1. a kind of Rosa roxburghii Tratt polyphenol extract, it is characterised in that wherein polyphenol content is not less than 33%.
2. a kind of preparation method of Rosa roxburghii Tratt polyphenol extract according to claim 1, it is characterised in that the following steps are included:
Step 1: taking fruit of Grossularia burejensis Berger, drying and crushing is added a certain amount of ethanol water and is heated to required temperature after crossing 50 meshes
Degree, extracts extracting solution under certain ultrasonic power；
Step 2: taking extracting solution, solvent is recovered under reduced pressure, certain temperature concentrated by rotary evaporation obtains Rosa roxburghii Tratt polyphenol crude extract；
Step 3: taking Rosa roxburghii Tratt polyphenol crude extract, fill column, the thick polyphenol of certain density Rosa roxburghii Tratt that will be made into ethyl alcohol with macroreticular resin
Solution, with column purification on certain sample solution concentration, sample solution flow rate and sample solution pH, then centainly to desorb flow velocity, solution
Imbibition concentration, stripping liquid pH afford eluent, and using concentration is dried under reduced pressure, vacuum drying obtains Rosa roxburghii Tratt polyphenol extract
Powder.
3. a kind of preparation method of Rosa roxburghii Tratt polyphenol extract according to claim 2, it is characterised in that ethanol water in step 1
The volume fraction of ethyl alcohol is 0%-100% in solution；Quality-volume ratio of fruit of Grossularia burejensis Berger and the ethanol water is 1:5-
1:150；Heating temperature is 25 DEG C -100 DEG C；Ultrasonic power is 80-400W；The extraction time of extracting solution is 0.5h-4h；Extracting solution
Extraction time be 1-4 times.
4. a kind of preparation method of Rosa roxburghii Tratt polyphenol extract according to claim 2, it is characterised in that ethanol water in step 1
The volume fraction of ethyl alcohol is 80% in solution；Quality-volume ratio of fruit of Grossularia burejensis Berger and the ethanol water is 1:100；Add
Hot temperature is 90 DEG C；Ultrasonic power is 250W；The extraction time of extracting solution is 80min, 40min；The extraction time of extracting solution is 3
It is secondary.
5. a kind of preparation method of Rosa roxburghii Tratt polyphenol extract according to claim 3 or 4, it is characterised in that the tool of step 2
Body is accomplished by the extracting solution of acquisition in 40-60 DEG C of rotary evaporation, and the ethyl alcohol in the extracting solution that step 1 obtains is passed through in removal,
Obtain Rosa roxburghii Tratt polyphenol crude extract.
6. a kind of preparation method of Rosa roxburghii Tratt polyphenol extract according to claim 5, it is characterised in that step 3 tool
Body is accomplished by the Rosa roxburghii Tratt polyphenol crude extract for obtaining step 2, is adsorbed with certain sample solution concentration by macroporous absorbent resin,
Successively with the water of certain pH certain flow rate and low-concentration ethanol be eluted to it is colourless after, successively use 40-95% concentration ethanol gradient
Elution, collects the ethanol eluate of 40-95%.
7. a kind of preparation method of Rosa roxburghii Tratt polyphenol extract according to claim 6, it is characterised in that described in step 3
Macroreticular resin is middle polarity or low pole macroreticular resin, including ADS-17, HPD-600, AB-8 and NKA-9.
8. a kind of preparation method of Rosa roxburghii Tratt polyphenol extract according to claim 7, it is characterised in that described in step 3
Loading flow velocity is 1-5mL/min；Sample solution concentration is 0.6-1.0mg/mL；Sample solution pH value is 4-8.
9. a kind of preparation method of Rosa roxburghii Tratt polyphenol extract described according to claim 6 or 7 or 8, it is characterised in that in step 3
The desorption flow velocity be 1-5mL/min；Stripping liquid concentration is 50%-90%；Stripping liquid pH value is 4-8.
10. application of the Rosa roxburghii Tratt polyphenol extract according to claim 1 in cosmetics, it is characterised in that: Rosa roxburghii Tratt polyphenol
Extract can be as the raw material of skin-lightening cosmetic.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111297765A (en) * | 2020-04-14 | 2020-06-19 | 遵义医科大学珠海校区 | Roxburgh rose extract and antibacterial application thereof |
| CN114680252A (en) * | 2022-03-30 | 2022-07-01 | 中国农业大学 | Low-astringency transparent roxburgh rose oral liquid and preparation method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102353745A (en) * | 2011-08-26 | 2012-02-15 | 贵州师范大学 | Preparation method for roxburgh rose and multi-index detection method |
| CN107308033A (en) * | 2017-05-22 | 2017-11-03 | 福建省亚热带植物研究所 | A kind of preparation method of olive fruits high-purity polyphenol and its application in cosmetics |
| CN107362080A (en) * | 2017-07-18 | 2017-11-21 | 上海应用技术大学 | A kind of extracting method of polyphenol in passion fruit pericarp |
| CN109330934A (en) * | 2018-10-30 | 2019-02-15 | 天峨县科学技术情报研究所 | The method of apple polyphenol is extracted from apple |
| CN109864943A (en) * | 2019-04-03 | 2019-06-11 | 黄山峰源生物科技有限公司 | A kind of preparation method and application of bletilla striata polyphenol |
| CN109984975A (en) * | 2019-05-16 | 2019-07-09 | 江苏省中国科学院植物研究所 | With the Preparation method and use of antioxidant activity total phenol in a kind of apocarya grouts |
-
2018
- 2018-09-11 CN CN201811057674.XA patent/CN109091429A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102353745A (en) * | 2011-08-26 | 2012-02-15 | 贵州师范大学 | Preparation method for roxburgh rose and multi-index detection method |
| CN107308033A (en) * | 2017-05-22 | 2017-11-03 | 福建省亚热带植物研究所 | A kind of preparation method of olive fruits high-purity polyphenol and its application in cosmetics |
| CN107362080A (en) * | 2017-07-18 | 2017-11-21 | 上海应用技术大学 | A kind of extracting method of polyphenol in passion fruit pericarp |
| CN109330934A (en) * | 2018-10-30 | 2019-02-15 | 天峨县科学技术情报研究所 | The method of apple polyphenol is extracted from apple |
| CN109864943A (en) * | 2019-04-03 | 2019-06-11 | 黄山峰源生物科技有限公司 | A kind of preparation method and application of bletilla striata polyphenol |
| CN109984975A (en) * | 2019-05-16 | 2019-07-09 | 江苏省中国科学院植物研究所 | With the Preparation method and use of antioxidant activity total phenol in a kind of apocarya grouts |
Non-Patent Citations (1)
| Title |
|---|
| 孙红艳: "《超声波法提取刺梨多酚的工艺优化及体外抑菌活性研究》", 《中国食品添加剂》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111297765A (en) * | 2020-04-14 | 2020-06-19 | 遵义医科大学珠海校区 | Roxburgh rose extract and antibacterial application thereof |
| CN114680252A (en) * | 2022-03-30 | 2022-07-01 | 中国农业大学 | Low-astringency transparent roxburgh rose oral liquid and preparation method thereof |
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