CN115054551B

CN115054551B - A composition containing radix Angelicae sinensis, atractylodis rhizoma and silk with speckle removing and antioxidant effects and its cosmetic application

Info

Publication number: CN115054551B
Application number: CN202210815196.4A
Authority: CN
Inventors: 钟姝凝
Original assignee: Zaozuo Technology Co ltd
Current assignee: Zaozuo Technology Co ltd
Priority date: 2022-07-09
Filing date: 2022-07-09
Publication date: 2023-05-12
Anticipated expiration: 2042-07-09
Also published as: CN115054551A

Abstract

The application discloses a freckle-removing and antioxidant modified composition containing angelica sinensis, bighead atractylodes rhizome and silk and application of the composition in cosmetics. The application fully plays the roles of relevant antioxidant, freckle removing and vascular active ingredient activating in angelica. The silk fibroin microsphere is prepared by utilizing silk fibroin in silk, angelica extract and white atractylodes rhizome powder are carried, the prepared nano microsphere can fully exert the synergistic effect of the related activities in the angelica and white atractylodes rhizome powder, and cell experiments and animal experiments prove that the nano microsphere has the effects of resisting oxidization, inhibiting tyrosinase, reducing melanin deposition and promoting blood circulation by removing blood stasis. The cosmetic prepared by taking the extract as a raw material not only beautifies skin and removes freckles, but also can resist oxidation, activate blood and remove stasis, and has good market prospect.

Description

A composition containing radix Angelicae sinensis, atractylodis rhizoma and silk with speckle removing and antioxidant effects and its cosmetic application

Technical Field

The invention belongs to the field of cosmetic, and particularly relates to a skin care product, in particular to a freckle-removing and oxidation-resisting modified composition containing angelica sinensis and silk and application of the composition in cosmetics.

Background

The macula is macula formed by melanin deposited on human skin, and comprises freckle, black speck, chloasma, sunburn, radiation spot, senile plaque, etc., and belongs to dyschromatosis skin diseases. The color spots are caused by the combined action of various factors, and can cause the activation of tyrosinase pathways in melanocytes and the excessive synthesis of melanin due to the reasons of long-term sun exposure, disorder of hormone levels in bodies, excessive pressure, poor quality cosmetics and the like.

Dang Gui is sweet and pungent in nature and warm in nature. It enters liver, heart and spleen meridians. Has effects of replenishing blood, promoting blood circulation, regulating menstruation, relieving pain, and loosening bowel to relieve constipation. The materia medica outline of the invention records that angelica can treat headache, pain in heart and abdomen, moisten the skin of the intestine and stomach bones and muscles, treat carbuncle, expel pus and relieve pain, and regulate blood and enrich blood. The angelica contains a large amount of organic components such as ligustilide, ferulic acid, chlorogenic acid and the like, and the ligustilide has the functions of relaxing smooth muscle, inhibiting bacteria, improving the body immunity regulation and the like; ferulic acid has radioprotective, antioxidant, antiviral, and antibacterial effects. The Atractylodis rhizoma contains active ingredients such as canker, mannan AM-3, caryophyllene, etc. The active ingredients in the bighead atractylodes rhizome enable the bighead atractylodes rhizome to have certain functions of resisting oxidation and aging, enhancing organism immunity and the like.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to prepare the cosmetic which has the characteristics of freckle removal, oxidation resistance, blood vessel activation and the like. In order to achieve the above object, the present invention is realized by the following technical scheme:

in a first aspect, the present invention provides a speckle-removing antioxidant modified composition comprising: angelica sinensis, bighead atractylodes rhizome and fibroin.

In a second aspect, the invention provides a preparation method of nano-microspheres of angelica sinensis and bighead atractylodes rhizome modified silk fibroin, which comprises the following steps:

s1, preparing an angelica extract: weighing 30-50g of radix Angelicae sinensis powder, adding methanol 20-30ml, weighing, performing ultrasonic treatment for 20-50min, adding methanol to balance weight, shaking, filtering, collecting filtrate, and drying to obtain radix Angelicae sinensis extract.

S2, preparing bighead atractylodes rhizome powder: taking 10-50g of bighead atractylodes rhizome, respectively adding 10 times of water, extracting for 2 times at 100 ℃,1 st time for 3h, 2 nd time for 2h, combining filtrates, concentrating under reduced pressure, vacuum drying, and crushing to obtain bighead atractylodes rhizome powder.

S3, preparation of degummed silk: dissolving 0.5g of Na2CO3 in 1L of ultrapure water, adding 5g of silk, stirring continuously at 100 ℃, taking out the silk after 30min, washing with the ultrapure water at 60 ℃ to remove sericin and residual ions, repeating the above operation for three times, and after degumming, loosening silk clusters, and drying in an oven at 45-60 ℃ to obtain degummed silk.

S4, silk fibroin: weighing degummed silk, placing into a centrifuge tube, adding lithium bromide solution, shaking vigorously for 1min, placing the solution into a water bath at 60 ℃ and heating for 30min to obtain transparent viscous silk fibroin solution, centrifuging the obtained silk fibroin solution in a centrifuge for 1min, and freezing and freeze-drying by dialysis.

S5, adding 1-5ml of angelica extract and white atractylodes rhizome powder solution dissolved in ethanol into the silk fibroin solution, incubating for 24-36h at a certain temperature, centrifuging the mixture, and further drying to obtain the angelica modified silk fibroin nanospheres.

The research team of the invention discovers that the white atractylodes rhizome, the angelica extract and the silk fibroin are prepared into the nano microsphere by self-assembly, so that the active ingredients of the white atractylodes rhizome and the angelica extract can be preserved, and the drug effect can be exerted for a longer time. And the nano-microspheres can be better absorbed by the skin, and after the nano-microspheres are absorbed, the bighead atractylodes rhizome and the Chinese angelica in the nano-microspheres can better enter the skin and better play a role. After the white atractylodes rhizome and the angelica sinensis extract are prepared into the nano microsphere, the oxidation resistance and the freckle removing capability are obviously improved, but the oxidation resistance and the freckle removing capability of the white atractylodes rhizome and the angelica sinensis water extract are far less than the effect of the nano microsphere.

The research team of the invention also finds that the prepared nano microsphere also has a certain function of activating capillary vessels, so that blood stasis in the blood vessels is effectively removed. A large number of experiments prove that after the white atractylodes rhizome, the Chinese angelica and the silk fibroin are prepared into the nano microsphere, the chemical components of the white atractylodes rhizome, the Chinese angelica and the silk fibroin can be changed to a certain extent, so that the nano microsphere has the effects of promoting blood circulation and removing blood stasis.

Compared with the prior art, the invention has the following beneficial effects:

1. the nano microsphere in the invention has no hormone, pigment, synthetic spice, preservative and other harmful chemical additives.

2. The angelica and the bighead atractylodes rhizome have the effect of inhibiting the aminopeptidase, so that the effects of reducing the speckles and removing the speckles are achieved.

3. The modified composition disclosed by the invention has the advantages that the angelica and the bighead atractylodes rhizome are wrapped in the silk fibroin nano-microsphere, so that a synergistic effect can be achieved, and the nano-microsphere has higher antioxidation and freckle removing performances than those of the angelica or bighead atractylodes rhizome with a single component and has a good repairing effect on skin.

4. The invention has simple operation and lower production cost, and is beneficial to mass production.

5. The nano microsphere prepared by the invention also has the functions of activating capillary vessels and removing blood stasis.

Drawings

FIG. 1 is a scan of a nanoparticle of example 1 prepared according to the present invention.

FIG. 2 is a scan of a nanoparticle of example 3 prepared in accordance with the present invention.

FIG. 3 is a bar graph of DPPH clearance for the microspheres of examples 1-5 and comparative examples 1-3 prepared according to the present invention.

FIG. 4 is a bar graph of ABTS clearance for example 1-5 and comparative example 1-3 nanomicrospheres prepared according to the present invention.

FIG. 5 is a bar graph of cell viability for examples 1-5 and comparative examples 1-3 prepared according to the present invention.

FIG. 6 is a histogram of melanin content measurement of examples 1-5 and comparative examples 1-3 prepared according to the present invention.

Detailed Description

Other advantages and effects of the present invention will become apparent to those skilled in the art from the following disclosure, which describes the embodiments of the present invention with reference to specific examples. The invention may be practiced or carried out in other embodiments that depart from the specific details, and the details of the present description may be modified or varied from the spirit and scope of the present invention. It should be noted that the following embodiments and features in the embodiments may be combined with each other without conflict.

Preparation of silk fibroin nano microsphere carrying angelica sinensis and bighead atractylodes rhizome

1. Materials and methods

1. Preparation of silk fibroin nano microsphere carrying angelica sinensis and bighead atractylodes rhizome

The preparation process of the nano microsphere carrying the Chinese angelica and the bighead atractylodes rhizome silk fibroin of the specific embodiment 1 is as follows:

s1, preparing an angelica extract: weighing 30g of radix Angelicae sinensis powder, adding methanol 20ml, weighing, performing ultrasonic treatment for 20min, adding methanol to balance weight, shaking, filtering, collecting filtrate, and drying to obtain radix Angelicae sinensis extract.

S2, preparing bighead atractylodes rhizome powder: taking 10g of bighead atractylodes rhizome, respectively adding 10 times of water, extracting for 2 times at 100 ℃,1 st time for 3 hours and 2 nd time for 2 hours, combining the filtrates, concentrating under reduced pressure, vacuum drying and crushing to obtain bighead atractylodes rhizome powder.

S3, preparation of degummed silk: 5g of Na ₂ CO ₃ Dissolving in 1L ultrapure water, adding 7g silk, and heating at 100deg.CContinuously stirring, taking out silk after 30min, washing with 60 ℃ ultrapure water to remove sericin and residual ions, repeating the above operation for three times, pulling loose silk clusters after degumming, and drying in a drying oven at 45 ℃ to obtain degummed silk.

S4, silk fibroin: weighing degummed silk, placing into a centrifuge tube, adding 9.3mol/L lithium bromide solution, shaking vigorously for 1min, placing the solution into a water bath at 60 ℃ for heating for 30min to obtain transparent viscous silk fibroin solution, centrifuging the obtained silk fibroin solution in a centrifuge for 1min, and freezing, dialyzing and freeze-drying.

S5, rapidly injecting ethanol into a 3% silk fibroin solution according to the volume ratio of 1:4, performing ultrasonic treatment for 2min at low power by adopting an ultrasonic cell disruption instrument, performing self-assembly for 10min at room temperature, putting the solution at-20 ℃ for thorough freezing for 24h, taking out, slowly thawing the solution at room temperature to form a milky solution, centrifuging the emulsion at 9000r/min for 10min at 4 ℃ and performing freeze drying for later use. 1ml of angelica extract (1 mg/ml) dissolved in ethanol and white atractylodes rhizome powder (1 mg/ml) solution are added into 25mg of silk fibroin solution, and incubated for 24 hours at 30 ℃ and 160r/min, the mixture is centrifuged, and the mixture is further dried to obtain the angelica modified silk fibroin nano-microsphere.

The preparation process of the nano microsphere carrying the angelica sinensis and the bighead atractylodes rhizome silk fibroin in the specific embodiment 2 is approximately the same as that in the embodiment 1, and the difference is that the concentration of the angelica sinensis extract in the step S5 is 3mg/ml and the concentration of the bighead atractylodes rhizome powder is 1mg/ml.

The preparation process of the nano microsphere carrying the angelica sinensis and the bighead atractylodes rhizome silk fibroin in the specific embodiment 3 is approximately the same as that in the embodiment 1, and the difference is that the concentration of the angelica sinensis extract in the step S5 is 1mg/ml and the concentration of the bighead atractylodes rhizome powder is 3mg/ml.

The preparation process of the nano microsphere carrying the angelica sinensis and the bighead atractylodes rhizome silk fibroin of the specific embodiment 4 is approximately the same as that of the embodiment 1, and the difference is that the concentration of the angelica sinensis extract in the step S5 is 3mg/ml and the concentration of the bighead atractylodes rhizome powder is 3mg/ml.

The preparation process of the nano microsphere carrying the angelica sinensis and the bighead atractylodes rhizome silk fibroin in the specific embodiment 5 is approximately the same as that in the embodiment 1, and the difference is that the concentration of the angelica sinensis extract in the step S5 is 5mg/ml and the concentration of the bighead atractylodes rhizome powder is 5mg/ml.

The preparation process of the specific silk fibroin nanoparticle of comparative example 1 carrying radix Angelicae sinensis and Atractylodis rhizoma is substantially the same as that of example 1, except that radix Angelicae sinensis extract and Atractylodis rhizoma powder are not added in step S5.

The preparation process of the specific silk fibroin nanoparticle of comparative example 2 carrying angelica sinensis and bighead atractylodes rhizome is substantially the same as that of example 1, except that only angelica sinensis is added in step S5, and bighead atractylodes rhizome powder is not added.

The preparation process of a specific nano microsphere carrying angelica sinensis and bighead atractylodes rhizome silk fibroin in comparative example 3 is approximately the same as that of example 1, except that only bighead atractylodes rhizome powder is added in step S5, and angelica sinensis is not added.

The preparation process of the nano microsphere carrying the Chinese angelica and the bighead atractylodes rhizome silk fibroin of the specific embodiment 6 is as follows:

s1, preparing an angelica extract: weighing 50g of radix Angelicae sinensis powder, adding 20ml of methanol, weighing, performing ultrasonic treatment for 20min, adding methanol to balance weight, shaking, filtering, collecting filtrate, and drying to obtain radix Angelicae sinensis extract.

S2, preparing bighead atractylodes rhizome powder: extracting 30g Atractylodis rhizoma with 10 times of water at 100deg.C for 2 times (1 st time and 3h, 2 nd time and 2 h), mixing filtrates, concentrating under reduced pressure, vacuum drying, and pulverizing to obtain Atractylodis rhizoma powder.

S3, preparation of degummed silk: 3g of Na ₂ CO ₃ Dissolving in 1L of ultrapure water, adding 6g of silk, stirring continuously at 100deg.C, taking out silk after 30min, washing with 60 ℃ ultrapure water to remove sericin and residual ions, repeating the above operation for three times, loosening silk cluster after degumming, and oven drying at 45deg.C to obtain degummed silk.

The preparation process of a specific nano microsphere of silk fibroin of comparative example 4 carrying angelica and bighead atractylodes rhizome is substantially the same as that of example 6, and the difference is that: s1, preparing an angelica extract: adding 50ml water into 30g radix Angelicae sinensis powder, extracting for 2 hr for the first time, extracting for 1 hr for the second time, extracting the mixed solution, filtering, concentrating, and drying and dehydrating to get radix Angelicae sinensis extract.

2. Measurement of antioxidant Properties

(1) DPPH radical scavenging Capacity determination

The results are expressed as IC50 by linear regression analysis of dose response curves plotted between DPPH radical scavenging performance and extract concentration. IC50 refers to the mass concentration (mg/ml) of the extract required to reduce the DPPH radical concentration by 50%, the lower the IC50 value, the higher the radical scavenging activity.

The absorbance (1.+ -. 0.02) of 6ml of DPPH methanol solution was mixed with the samples of experimental examples 1 to 5 and comparative examples 1 to 3. The reaction mixture was shaken at room temperature, allowed to stand in the dark for 20min, and the change in absorbance value was measured at a wavelength of 517 nm. The scavenging performance of the solution of the extract sample replaced by methanol as a blank control sample on DPPH free radicals is calculated according to the formula:

DPPH radical scavenging ability/% = (a) _{Control group} -A _{Sample group} )/A _{Control group} ×100

(2) ABTS radical scavenging ability assay

Dissolving ABTS in 20mmol/L acetic acid buffer solution with pH of 4.5 to obtain 7mmol/L ABTS cation free radical stock solution, mixing with potassium persulfate solution (final concentration of 2.45mmol/L in the mixed system), and reacting at room temperature in dark for 16h to make the solution reach stable oxidation state. The ABTS cationic free radical working solution is obtained by diluting the ABTS free radical stock solution with 20mmol/L acetic acid buffer solution with pH of 4.5 according to a proper proportion (volume ratio of 1:75) before measurement to enable the absorbance value of the solution to reach (0.7+/-0.02) at the wavelength of 734 nm. The reaction system at the time of measurement included 3ml of ABTS cationic radical working solution and samples of experimental examples 1-5 and comparative examples 1-3. After standing for 10min at room temperature in dark, measuring the absorbance at 734nm wavelength, taking methanol instead of the extract sample solution as blank control, and calculating the scavenging performance of the sample on ABTS cation free radicals according to the formula:

ABTS radical scavenging ability% = (a _{Control group} -A _{Sample group} )/A _{Control group} ×100

The results are expressed as IC50 by linear regression analysis of dose response curves plotted between ABTS cation radical scavenging performance and extract concentration.

3. Tyrosinase inhibition activity assay

In skin melanin biosynthesis tyrosinase is mostly used for dopa to form dopaquinone, which forms melanin after a series of spontaneous reactions.

Preparing reaction solution according to the table 1, precisely transferring test samples, respectively adding into 96-well plates, uniformly mixing, incubating at 37 ℃ for 10min, rapidly adding 100 mu L of dopa solution, and uniformly mixing. The OD value of the solution AT 475nm was measured continuously for 30min with phosphate buffered saline PBS as a control to obtain AC1, AC2, AT1 and AT2, which were measured in parallel for 3 times. The tyrosinase inhibitory activity of the samples was calculated as follows:

tyrosinase activity inhibition ratio= [1- (a) _T1 -A _T2 )/(A _C1 -A _C2 )]×100％

TABLE 1

2. Results

TABLE 2

As is clear from Table 2, the DPPH radical scavenger, ABTS radical scavenger, tyrosinase inhibition rate, etc. of examples 1-5 were significantly lower than those of comparative examples 1-3, indicating that the antioxidant properties of examples 1-5 were higher than those of comparative examples. In addition, the DPPH free radical inhibition rate, the ABTS free radical removal rate and the tyrosinase inhibition rate of the sample all show a decreasing trend along with the increase of the contents of the angelica and the white atractylodes rhizome, which indicates that the oxidation resistance of the sample is improved along with the increase of the contents of the white atractylodes rhizome and the angelica. And the embodiment of adding the bighead atractylodes rhizome and the Chinese angelica at the same time is higher than the embodiment of adding the single content of the comparative example, which shows that the common addition of the bighead atractylodes rhizome and the Chinese angelica has a synergistic effect, so that the oxidation resistance is obviously improved.

Fig. 1 and fig. 2 are scanned views of the nano-microspheres prepared in the embodiment 1 and the embodiment 3 of the present invention, and it can be seen from the figures that the diameter of the nano-microsphere is 300-500nm, the size of the nano-microsphere is basically consistent, and the stability can be maintained higher.

FIG. 3 is a bar chart of DPPH clearance and FIG. 4 is a bar chart of ABTS clearance of the nano-microspheres prepared by the invention. It is apparent from the figures that examples 1 to 5 have excellent oxidation resistance.

TABLE 3 Table 3

	Ferulic acid extraction yield (%)	Ligustilide extraction yield (%)
			Example 6	547.8±5.6	250.9±7.2
Comparative example 4	427.5±4.8	247.7±6.3

The active ingredients in the angelica have strong volatility, mainly comprise phthalide compounds such as butenyl phthalide, ligustilide and the like, and the extraction process can lead to: the drug concentration of the extract of (1) cannot be maintained constant. (2) Ferulic acid has extremely poor stability in solution and can be decomposed more rapidly in a neutral solvent under the light of the environment. (3) Because the solubility of the ferulic acid in water is small, the stability is poor, and the ferulic acid can be seriously damaged in a high-temperature environment. As can be seen from Table 3, the contents of ferulic acid and ligustilide in the Angelica sinensis extract extracted by methanol diafiltration are significantly higher than those in the conventional water extraction. The extraction method used in the application is better than the traditional water extraction method.

Cell experiment

1. Materials and methods

1. Material

B16-F10 cells (Woheprunocel Life technologies Co., ltd.); south DMEM medium (Gibco, usa); 0.25% trypsin-EDTA (Gibco Co., U.S.A.); dimethyl sulfoxide (DMSO, sigma, usa); PBS buffer (Beijing Soy Bao technologies Co., ltd.); thiazole blue kit (MTT, jiangsu Kaiyi Co.).

2. Experiments for melanin inhibition

The melanin content in the cells was determined by NaOH cleavage. Digesting B16 cells in logarithmic phase, inoculating into 6-well plate at concentration of 2×10 ⁵ The cells/wells were incubated at 37℃with 5% CO2Culturing in incubator overnight. After the cells are completely attached, the cells are taken, and the culture is continued for 72 hours. The supernatant was discarded, washed 2 times with PBS, and 200. Mu.L of cell lysate was added to each well after cell scraping was used to collect the cells. After extensive lysis at 4℃for 40min, centrifugation was carried out at 12000rpm for 20min. The supernatant was transferred to an EP tube to determine protein concentration. After adding 190. Mu.L of 1mol/L NaOH (10% DMSO) lysate to the precipitate and fully dissolving in a water bath at 80℃for 1h, the absorbance was measured at 405nm and the final melanin content was normalized to the relative protein concentration. The calculation formula is as follows:

melanin relative content= (OD _{405 sample} Protein concentration)/(OD _{405 blank} Protein concentration) ×100%

Each sample was assayed in 3 replicates.

3. Cytotoxicity detection

Collecting cells in logarithmic growth phase, plating to obtain cells to be tested with density of 5000 cells/well, culturing at 37deg.C under 5% CO2, and growing cells to 10 ⁵ The whole DMEM medium containing 0.1mg/ml of the test solution was added to the cell suspension per ml of the cell suspension, and the same volume of the whole DMEM medium was added as a control group. The culture was continued for 24 hours, 20. Mu.L of 0.5% MTT solution (5 g/L) was added to each well, the culture solution was discarded after 4 hours of culture, and after 2-3 times of washing with PBS, 150. Mu.L of dimethyl sulfoxide was added to each well, and the mixture was shaken on a shaker at a low speed for 10 minutes to sufficiently dissolve the crystals. The absorbance of each well was measured at 490 nm.

Cell viability = (assay well absorbance-blank absorbance)/(control absorbance-blank absorbance) ×100%.

4. Blood circulation activating and stasis removing effect detection

(1) Acute blood stasis rat model establishment

Except blank groups, the rest experimental groups inject epinephrine hydrochloride injection 0.8mg.kg subcutaneously into rats ^-1 The total times are separated by 4 hours each time, so that a rat acute blood stasis model is formed.

(2) Grouping, administration and sampling

Rats were divided into a blank group, a model group, a positive control group, and an experimental group, and 0.9% NaCl20ml.kg was administered to the blank group and the model group ^-1 Positive pair73 mg/kg compound red sage root dripping pill for group administration ^-1 ·d ^-1 Each of the Chinese medicine administration groups was administered with 10.8 g.kg of the samples of example 1 and comparative examples 1 to 4 ^-1 ·d ^-1 The administration was performed by gastric lavage, 1 time a day, and 9 days after each other. After the 7 th day of gastric lavage, an acute blood stasis syndrome model was prepared. After molding, normal stomach irrigation is carried out on the 8 th day; after 30min of administration on day 9, 2ml of abdominal aorta was collected and added to an anticoagulant tube containing disodium citrate for measuring the blood coagulation index of 4.

2. Results

TABLE 4 Table 4

PT in the table is prothrombin time; APTT is the activated partial thromboplastin time; FIB is fibrinogen.

As can be seen from Table 4, the PT, APTT and FIB of the rat of example 1 are significantly lower than those of the blank group, indicating that example 1 has a certain effect of promoting blood circulation to remove blood stasis. As can be seen from the comparison of the example 1 and the comparative examples 1-3, the effect of the single component of angelica sinensis or bighead atractylodes rhizome on promoting blood circulation and removing blood stasis is very small, while the effect of the example 1 is remarkable, which shows that the addition of bighead atractylodes rhizome remarkably enhances the effect of angelica sinensis on promoting blood circulation and removing blood stasis, probably because the addition of bighead atractylodes rhizome combines part of active components of angelica sinensis and active components of bighead atractylodes rhizome, and the ingredient with remarkable effect of promoting blood circulation and removing blood stasis is generated.

FIG. 5 is a graph showing the intracellular melanin content of examples 1-5, comparative examples 1-3, and a blank. As can be seen from the figures, the melanin content of the cells in examples 1-5 was lower than that in comparative examples 1-3, indicating that the samples of examples 1-5 had some inhibitory effect on melanin production. With the increase of the contents of the angelica and the bighead atractylodes rhizome, the content liquid of melanin in cells gradually decreases, but the effect of the composition of the angelica and the bighead atractylodes rhizome is better than that of a single component.

As can be seen from table 4 and fig. 5, the effect of activating blood circulation to dissipate blood stasis and the effect of inhibiting melanin of a single component are not obvious, which indicates that unexpected changes occur in the nano-microsphere, and it is possible that the active ingredient in the bighead atractylodes rhizome reacts with the angelica and the silk fibroin in the nano-microsphere, so that the inhibiting rate of melanin is greatly improved, and in addition, a component is generated, so that the nano-microsphere has the effect of activating blood circulation to dissipate blood stasis.

FIG. 6 is a graph of cell viability of in vitro cell experiments examples 1-5 and comparative examples 1-3, and a blank. As can be seen, the cell viability was higher in examples 1-5 than in the blank, comparative examples 1-3, indicating that the samples of examples were non-toxic to the cells.

In conclusion, the freckle-removing and antioxidant composition prepared by the invention takes the angelica sinensis extract and the bighead atractylodes rhizome extract as raw materials, and is wrapped in the nano microsphere prepared by the silk fibroin, so that the angelica sinensis extract and the bighead atractylodes rhizome extract are better absorbed by a human body. The Chinese angelica and the largehead atractylodes rhizome can play roles in protecting a user from radiation, resisting oxidation, removing freckles and the like, and meanwhile, the mixture of the Chinese angelica and the largehead atractylodes rhizome can ensure that the ferulic acid plays a larger role, and plays roles in promoting blood circulation to remove blood stasis and relieving blood vessels. The method provides a new thought for the freckle-removing and antioxidant cosmetics and a new method for preparing the freckle-removing and antioxidant composition.

Finally, it should be noted that: the foregoing examples merely illustrate specific embodiments of the invention, which are described in greater detail and are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention.

Claims

1. A preparation method of a freckle-removing and oxidation-resisting modified composition containing angelica sinensis, bighead atractylodes rhizome and silk is characterized by comprising the following steps:

s1, preparing an angelica extract: weighing 30-50g of radix Angelicae sinensis powder, adding methanol 20-30ml, weighing, performing ultrasonic treatment for 20-50min, adding methanol to balance weight, shaking, filtering, collecting filtrate, and drying to obtain radix Angelicae sinensis extract;

s2, preparing bighead atractylodes rhizome powder: taking 10-50g of bighead atractylodes rhizome, respectively adding 10 times of water, extracting for 2 times at 100 ℃, carrying out 1 st time for 3h and 2 nd time for 2h, combining filtrates, concentrating under reduced pressure, vacuum drying, and crushing to obtain bighead atractylodes rhizome powder;

s3, preparation of degummed silk: dissolving 0.5g of Na2CO3 in 1L of ultrapure water, adding 5g of silk, stirring continuously at 100 ℃, taking out the silk after 30min, washing with the ultrapure water at 60 ℃ to remove sericin and residual ions, repeating the above operation for three times, and after degumming, loosening silk clusters, and drying in an oven at 45-60 ℃ to obtain degummed silk;

s4, silk fibroin: weighing degummed silk, placing into a centrifuge tube, adding lithium bromide solution, shaking vigorously for 1min, placing the solution into a water bath at 60 ℃ and heating for 30min to obtain transparent viscous silk fibroin solution, centrifuging the obtained silk fibroin solution in a centrifuge for 1min, and freeze-drying by dialysis;

s5, adding 1-5ml of angelica extract and white atractylodes rhizome powder solution dissolved in ethanol into the silk fibroin solution, incubating for 24-36h at a certain temperature, centrifuging the mixture, and further drying to obtain angelica modified silk fibroin nano microspheres;

the insoluble substances and bubbles in the solution are removed in the step S4, and the solution is put into a dialysis bag and dialyzed by ultrapure water for three days to completely remove the Li in the solution ⁺ And Br (Br) ^— Obtaining pure silk fibroin solution, pre-freezing the silk fibroin solution for 12 hours at the temperature of minus 80 ℃ in a refrigerator, and then putting the silk fibroin solution into a freeze dryer for freeze drying for 48 hours to obtain silk fibroin fibers.

2. The method of manufacturing according to claim 1, characterized in that: the concentration of the lithium bromide solution in the step S4 is 9.3mol/L.

3. The method of manufacturing according to claim 1, characterized in that: in the step S5, ethanol is rapidly injected into a 3% silk fibroin solution according to the volume ratio of 1:4, an ultrasonic cell disruption instrument is adopted to carry out ultrasonic treatment at low power, then the reaction is carried out at room temperature, the solution is placed at-20 ℃ to be thoroughly frozen for 24 hours and then taken out, the solution is slowly melted into a milky solution at room temperature, and the emulsion is centrifuged at 4 ℃ and then is frozen and dried for later use.

4. The method of manufacturing according to claim 1, characterized in that: the solution was incubated at 30℃for 24h at 160r/min in step S5.

5. The spot-removing antioxidant modified composition containing angelica sinensis, bighead atractylodes rhizome and silk prepared by the preparation method of any one of claims 1-4.

6. Use of the spot-removing antioxidant modified composition prepared by the preparation method according to claims 2-4 in the field of cosmetics.