CN115054551A

CN115054551A - Freckle-removing antioxidant modified composition containing angelica sinensis, bighead atractylodes rhizome and silk and application of freckle-removing antioxidant modified composition in cosmetics

Info

Publication number: CN115054551A
Application number: CN202210815196.4A
Authority: CN
Inventors: 钟姝凝
Original assignee: Hebei Xuande Biotechnology Co ltd
Current assignee: Zaozuo Technology Co ltd
Priority date: 2022-07-09
Filing date: 2022-07-09
Publication date: 2022-09-16
Anticipated expiration: 2042-07-09
Also published as: CN115054551B

Abstract

The application discloses a freckle-removing antioxidant modified composition containing Chinese angelica, bighead atractylodes rhizome and silk and application of the composition in the aspect of cosmetics. The application gives full play to the functions of relevant antioxidant, freckle removing and active components of activated blood vessels in the angelica. The silk fibroin microspheres are prepared from silk fibroin, and are loaded with angelica sinensis extract and bighead atractylodes rhizome powder, the prepared nano microspheres can fully play the synergistic effect of related activities in the angelica sinensis and the bighead atractylodes rhizome powder, cell experiments and animal experiments prove that the nano microspheres have an antioxidation effect, can inhibit the generation of tyrosinase and reduce the deposition of melanin, and meanwhile, the addition of the bighead atractylodes rhizome powder enables the nano microspheres to have a certain effect of promoting blood circulation and removing blood stasis. The cosmetics prepared by using the raw materials not only can beautify skin and remove freckles, but also can resist oxidation, promote blood circulation and remove blood stasis, and have good market prospect.

Description

Freckle-removing antioxidant modified composition containing angelica sinensis, bighead atractylodes rhizome and silk and application of freckle-removing antioxidant modified composition in cosmetics

Technical Field

The invention belongs to the field of makeup, and particularly relates to a skin care product, in particular to a freckle-removing antioxidant modified composition containing Chinese angelica and silk and application of the composition in the aspect of cosmetics.

Background

The stain is formed by deposition of melanin on human skin, and comprises freckle, black spot, chloasma, sunburn, radiation spot, senile plaque, etc., and belongs to pigmentation disorder dermatosis. The color spots are caused by the combined action of various factors, and the tyrosinase pathway in the melanoblast is activated due to long-term sun exposure, in-vivo hormone level imbalance, over-pressure, poor-quality cosmetic manufacturing and the like, so that the melanin is excessively synthesized.

Dang Gui is sweet, pungent and warm in nature. It enters liver, heart and spleen meridians. Has the functions of enriching blood, promoting blood circulation, regulating menstruation, relieving pain, moistening intestine and relaxing bowels. According to the records of the compendium of materia medica, Chinese angelica is used for treating headache, pain of heart and abdomen, moistening intestines, stomach, muscles and bones and skin, treating carbuncle-abscess, expelling pus, relieving pain, and tonifying 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 muscles, inhibiting bacteria, improving the body immunity and the like; ferulic acid has radioprotective, antioxidant, antiviral, and antibacterial effects. The Atractylodis rhizoma contains canodoxadone, mannan AM-3, caryophyllene, etc. as active ingredients. The active ingredients in the white atractylodes rhizome enable the white atractylodes rhizome to have certain functions of resisting oxidation and aging, enhancing the immunity of the organism 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 removing freckles, resisting oxidation, activating blood vessels and the like. In order to achieve the purpose, the invention is realized by the following technical scheme:

in a first aspect, the present invention provides a spot-removing antioxidant modification composition, which comprises: angelica, atractylodes and fibroin.

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

s1, preparing the angelica sinensis extract: weighing 30-50g of radix Angelicae sinensis powder, adding 20-30ml of methanol, weighing, ultrasonically treating for 20-50min, adding methanol to make up the weight, shaking, filtering, collecting filtrate, and drying to obtain radix Angelicae sinensis extract.

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

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

S4, silk fibroin: weighing the degumming silk, putting the degumming silk into a centrifugal tube, adding a lithium bromide solution, violently shaking for 1min, putting the solution into a water bath at 60 ℃ for heating for 30min to obtain a transparent viscous silk fibroin solution, centrifuging the obtained silk fibroin solution in a centrifugal machine for 1min, freezing, dialyzing, and freeze-drying.

S5, adding 1-5ml of angelica sinensis extract and bighead 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 sinensis modified silk fibroin nanospheres.

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

The research team of the invention also finds that the prepared nano-microspheres also have a certain effect of activating capillary vessels, so that the stasis in the blood vessels is effectively removed. A large number of experiments prove that after the bighead atractylodes rhizome, the Chinese angelica and the silk fibroin are prepared into the nano microspheres, chemical components of the bighead atractylodes rhizome, the Chinese angelica and the silk fibroin are changed to a certain extent, so that the nano microspheres have certain effects of activating blood and dissolving stasis.

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

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

2. The angelica and the bighead atractylodes rhizome in the invention have the function of inhibiting the tyrosinase, and achieve the effects of fading the scars and removing the freckles.

3. The modified composition of the invention wraps the angelica and the white atractylodes rhizome in the fibroin protein nano microspheres to achieve synergistic effect, and the nano microspheres have higher oxidation resistance and spot removing performance than the angelica or the white atractylodes rhizome with single component, and have good repairing effect on skin.

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

5. The nano-microspheres prepared by the invention also have certain effects of activating capillary vessels and removing blood stasis.

Drawings

FIG. 1 is a scanned image of the nanospheres of example 1 prepared in accordance with the present invention.

FIG. 2 is a scanned image of the nanospheres of example 3 prepared in accordance with the present invention.

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

FIG. 4 is a bar graph of ABTS clearance of the inventive and comparative examples 1-3 nanospheres.

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

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

Detailed Description

The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention. It is to be noted that the features in the following embodiments and examples may be combined with each other without conflict.

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

Materials and methods

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

A specific preparation process of the silk fibroin nanospheres carrying angelica and bighead atractylodes rhizome in example 1 is as follows:

s1, preparing the angelica sinensis extract: weighing radix Angelicae sinensis powder 30g, adding methanol 20ml, weighing, ultrasonic processing for 20min, adding methanol to make up weight, shaking, filtering, collecting filtrate, and drying to obtain radix Angelicae sinensis extract.

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

S3, preparing the degumming silk: adding 5g of Na ₂ CO ₃ Dissolving in 1L of ultrapure water, adding 7g of silk, stirring continuously at 100 ℃, taking out the silk after 30min, washing the silk with ultrapure water at 60 ℃ to remove sericin and residual ions, repeating the operation for three times, loosening the silk cluster after degumming, and drying in an oven at 45 ℃ to obtain the degummed silk.

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

S5, quickly injecting ethanol into 3% silk fibroin solution at a volume ratio of 1: 4, carrying out ultrasonic treatment for 2min at low power by using an ultrasonic cell disruptor, carrying out self-assembly for 10min at room temperature, completely freezing the solution at-20 ℃ for 24h, taking out, slowly melting the solution at room temperature to form milky solution, centrifuging the emulsion at 4 ℃ for 10min at 9000r/min, and freeze-drying for later use. Adding 1ml of radix Angelicae sinensis extract (1mg/ml) and Atractylodis rhizoma powder (1mg/ml) dissolved in ethanol into 25mg of silk fibroin solution, incubating at 30 deg.C and 160r/min for 24 hr, centrifuging the mixture, and further drying to obtain radix Angelicae sinensis modified silk fibroin nanospheres.

A specific process for preparing silk fibroin nanospheres carrying angelica and atractylodes macrocephala in example 2 is substantially the same as that in example 1, except that the concentration of the angelica extract is 3mg/ml and the concentration of the atractylodes macrocephala powder is 1mg/ml in step S5.

A specific process for preparing silk fibroin nanospheres carrying angelica and atractylodes macrocephala in example 3 is substantially the same as that in example 1, except that the concentration of the angelica extract is 1mg/ml and the concentration of the atractylodes macrocephala powder is 3mg/ml in step S5.

A specific process for preparing silk fibroin nanospheres carrying angelica and atractylodes macrocephala in example 4 is substantially the same as that in example 1, except that the concentration of the angelica extract is 3mg/ml and the concentration of the atractylodes macrocephala powder is 3mg/ml in step S5.

A specific process for preparing silk fibroin nanospheres carrying angelica and atractylodes macrocephala in example 5 is substantially the same as that in example 1, except that the concentration of the angelica extract is 5mg/ml and the concentration of the atractylodes macrocephala powder is 5mg/ml in step S5.

A specific preparation process of the silk fibroin nanospheres carrying angelica and white atractylodes rhizome of comparative example 1 is substantially the same as that of example 1, except that no angelica extract and no white atractylodes rhizome powder are added in step S5.

A specific preparation process of the silk fibroin nanospheres carrying angelica and white atractylodes rhizome in comparative example 2 is substantially the same as that in example 1, except that only angelica is added in step S5, and no white atractylodes rhizome powder is added.

A specific preparation process of the silk fibroin nanospheres carrying angelica and atractylodes macrocephala of comparative example 3 is substantially the same as that of example 1, except that only the powder of atractylodes macrocephala is added in step S5, and no angelica is added.

A specific process for preparing the silk fibroin nanospheres carrying angelica and atractylodes macrocephala of example 6 is as follows:

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

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

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

A specific comparative example 4 is substantially the same as example 6 in the preparation process of the silk fibroin nanospheres carrying angelica and atractylodes macrocephala, except that: s1, preparing the angelica sinensis extract: adding 50ml water into 30g radix Angelicae sinensis powder, extracting for the first time for 2 hr and the second time for 1 hr, extracting the mixed solution, filtering, concentrating, and drying and dehydrating to obtain radix Angelicae sinensis extract.

2. Measurement of antioxidant Property

(1) DPPH free radical scavenging ability measurement

The results were expressed as IC50 by linear regression analysis of the dose-effect curve plotted between DPPH radical scavenging performance and extract concentration. IC50 refers to the mass concentration value (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.

6ml of DPPH methanol solution absorbance (1. + -. 0.02) was mixed with the samples of Experimental examples 1-5 and comparative examples 1-3. The reaction mixture was shaken up at room temperature, left to stand in the dark for 20min, and the change in absorbance at a wavelength of 517nm was measured. The DPPH free radical scavenging performance of a blank control sample which is a sample solution of methanol instead of an extract is calculated according to the formula:

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

(2) ABTS free radical scavenging Capacity assay

ABTS is dissolved in acetic acid buffer solution with the concentration of 20mmol/L and the pH value of 4.5 to obtain ABTS cation free radical liquid with the concentration of 7mmol/L, and the ABTS cation free radical liquid is mixed with potassium persulfate solution (the final concentration in a mixed system is 2.45mmol/L) and reacted for 16 hours in a dark place at room temperature to enable the solution to reach a stable oxidation state. Before measurement, ABTS free radical stock solution is diluted by 20mmol/L acetic acid buffer solution with pH4.5 according to a proper proportion (volume ratio is 1:75) to make the absorbance value of the solution at 734nm reach (0.7 +/-0.02), thereby obtaining ABTS cationic free radical working solution. The reaction system comprises 3ml of ABTS cationic free radical working solution and samples of experimental examples 1-5 and comparative examples 1-3. Standing at room temperature in dark place for 10min, measuring light absorption value at 734nm wavelength, using methanol to replace extract sample solution as blank control, and calculating ABTS cation free radical scavenging performance of the sample according to formula:

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

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

3. Tyrosinase inhibitory Activity assay

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

Reaction solutions were prepared as shown in Table 1, and the samples were precisely transferred and added to 96-well plates, mixed well, incubated at 37 ℃ for 10min, then 100. mu.L of dopa solution was rapidly added, and mixed well. The OD value of the solution AT 475nm was measured continuously for 30min using PBS as a control, to obtain AC1, AC2, AT1 and AT2, which were measured in parallel 3 times. The tyrosinase inhibiting activity of the sample was calculated according to the formula:

tyrosinase activity inhibition rate ═ 1- (A) _T1 -A _T2 )/(A _C1 -A _C2 )]×100％

TABLE 1

Second, result in

TABLE 2

As can be seen from Table 2, the DPPH radical clearance, ABTS radical clearance, tyrosinase inhibition, and the like of examples 1-5 are all significantly lower than those of comparative examples 1-3, indicating that the antioxidant performance of examples 1-5 is higher than that of comparative examples. In addition, with the increase of the contents of angelica and white atractylodes rhizome, the DPPH free radical inhibition rate, ABTS free radical clearance rate and tyrosinase inhibition rate of the sample show a trend of decreasing, which shows that with the increase of the contents of white atractylodes rhizome and angelica, the antioxidant performance of the sample is improved. The embodiment of adding the white atractylodes rhizome and the angelica is higher than the comparative example of adding the white atractylodes rhizome and the angelica with single content, which shows that the white atractylodes rhizome and the angelica have synergistic effect and obviously improve the oxidation resistance.

Fig. 1 and fig. 2 are scanned images of the nanospheres prepared in examples 1 and 3 of the present invention, and it can be seen from the images that the diameter of the nanosphere is between 300 and 500nm, the size of the nanosphere is substantially consistent, and the nanosphere can maintain high stability.

Fig. 3 and 4 are bar graphs of DPPH clearance and ABTS clearance of the nano-microspheres prepared by the invention. It is apparent from the figures that examples 1 to 5 have excellent antioxidant properties.

TABLE 3

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

When the effective components in the Chinese angelica have stronger volatility and are mainly phthalides such as butenyl phthalide, ligustilide and the like, the extraction process can cause that: (1) the drug concentration of the extract cannot be maintained constant. (2) Ferulic acid has very poor stability in solution and undergoes faster decomposition under light in a neutral solvent. (3) The solubility of ferulic acid in water is low, 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 content of ferulic acid and ligustilide in the Angelica sinensis extract obtained by methanol diafiltration was significantly higher than that obtained by conventional water extraction. It is shown that the extraction method used in this application is better than the traditional water extraction method.

Cell experiments

Materials and methods

1. Material

B16-F10 cells (Wuhan Punuo Sai Life technologies, Inc.); south DMEM medium (Gibco, usa); 0.25% trypsin-EDTA (Gibco, USA); dimethyl sulfoxide (DMSO, Sigma, usa); PBS buffer (beijing solibao science and technology ltd); thiazole blue kit (MTT, Kyoki Co., Ltd.).

2. Experiment for melanin inhibition

And (3) measuring the melanin content in the cells by adopting a NaOH cracking method. B16 cells in logarithmic growth phase were digested and seeded in 6-well plates at a concentration of 2X 10 ⁵ One well per well, and incubated overnight in a 5% CO2 incubator at 37 ℃. After the cells are completely attached to the wall, the medicine is taken and the culture is continued for 72 h. The supernatant was discarded, washed 2 times with PBS, and 200. mu.L of cell lysate was added to each well after collecting cells using a cell scraper. After lysis was carried out sufficiently at 4 ℃ for 40min, the cells were centrifuged at 12000rpm for 20 min. Transfer supernatant to EP tube for protein concentration. 190 mu.L of 1mol/L NaOH (containing 10% DMSO) lysate is added into the precipitate, the precipitate is fully dissolved in water bath at 80 ℃ for 1h, the absorbance value is measured at 405nm, and the final melanin content is normalized by using the relative protein concentration. The calculation formula is as follows:

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

Each sample was assayed in 3 replicates.

3. Cytotoxicity assays

Collecting cells in logarithmic growth phase, plating to make cell density to be tested reach 5000/hole, culturing at 37 deg.C under 5% CO2, and growing to 10 ⁵ Cell suspension of one cell per mlThe solution was changed and DMEM complete medium containing 0.1mg/ml of the test solution was added, and the control group was DMEM complete medium added in the same volume. Continuing to culture for 24h, adding 20 μ L0.5% MTT solution (5g/L) into each well, culturing for 4h, discarding the culture solution, washing with PBS for 2-3 times, adding 150 μ L dimethyl sulfoxide into each well, and shaking on shaking table at low speed for 10min to dissolve the crystal completely. The absorbance of each well was measured at 490 nm.

The cell viability was (assay well absorbance-blank control absorbance)/(control absorbance-blank control absorbance) × 100%.

4. Detection of blood circulation promoting and blood stasis removing effects

(1) Establishment of acute blood stasis rat model

Except for the blank group, the other experimental groups injected the adrenaline hydrochloride injection 0.8 mg.kg into the rat subcutaneously ^-1 The acute blood stasis model of the rat is created by 4 hours of each time.

(2) Grouping, administering, sampling

Dividing the rats into blank group, model group, positive control group and experimental group, and administering 0.9% NaCl20 ml-kg to the blank group and the model group ^-1 73 mg/kg compound radix Salviae Miltiorrhizae drop pill for positive control group ^-1 ·d ^-1 In each of the groups to which the Chinese medicinal materials were administered, 10.8 g/kg of the sample of example 1 and comparative examples 1 to 4 was administered ^-1 ·d ^-1 The administration is carried out by gavage 1 time per day for 9 days. After the 7 th day of intragastric administration, an acute blood stasis model was prepared. After molding, normal intragastric administration is carried out on day 8; after administration for 30min on day 9, 2ml of blood is collected from abdominal aorta, and the obtained blood is added into an anticoagulation tube containing disodium citrate for measuring 4-item coagulation indexes.

Second, result in

TABLE 4

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

As can be seen from Table 4, the PT, APTT and FIB of the rats in example 1 are significantly reduced compared with the blank group, which indicates that example 1 has certain effect of promoting blood circulation and removing blood stasis. It can be seen from the comparison between example 1 and comparative examples 1-3 that the effect of blood circulation-promoting and blood stasis-removing of the angelica or the atractylodes macrocephala of a single component is very slight, while the effect of example 1 is significant, which indicates that the effect of blood circulation-promoting and blood stasis-removing of the angelica is significantly enhanced by the addition of the atractylodes macrocephala, probably because the addition of the atractylodes macrocephala combines part of the active components of the angelica and the active components of the atractylodes macrocephala to generate the components with significant effects of blood circulation-promoting and blood stasis-removing.

FIG. 5 is a graph showing the intracellular melanin contents of examples 1 to 5, comparative examples 1 to 3, and a blank control. As can be seen from the figure, the melanin content of the cells in examples 1-5 is lower than that in comparative examples 1-3, which shows that the samples in examples 1-5 have a certain inhibitory effect on the production of melanin. With the increase of the content of angelica and atractylodes, the content of melanin in cells is gradually reduced, but the effect of the composition of angelica and atractylodes is better than that of a single component.

As can be seen from table 4 and fig. 5, the effect of promoting blood circulation by removing blood stasis and the inhibitory rate of melanin of a single component are not significant, which indicates that unexpected changes occur in the nanospheres, and it is likely that the active ingredient in the atractylodes macrocephala reacts with angelica sinensis and silk fibroin in the nanospheres, so that the inhibitory rate of the atractylodes macrocephala on melanin is greatly improved, and a component is generated, so that the nanospheres have the effect of promoting blood circulation by removing blood stasis.

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

In conclusion, the freckle-removing and antioxidant composition prepared by the invention takes the angelica extract and the bighead atractylodes rhizome extract as raw materials, and the raw materials are wrapped in the nano microspheres prepared from the silk fibroin, so that the angelica extract and the bighead atractylodes rhizome extract are better absorbed by a human body. The angelica and the atractylodes have the effects of protecting a user from radiation, resisting oxidation, removing freckles and the like, and the ferulic acid plays a greater role by mixing the angelica and the atractylodes, and plays roles of promoting blood circulation to remove blood stasis and relieving blood vessels. The method provides a new idea 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 above-mentioned embodiments only express the specific embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention.

Claims

1. A freckle-removing antioxidant modified composition containing Chinese angelica, bighead atractylodes rhizome and silk is characterized by comprising the following components in percentage by weight:

(1) preparing extracts of rhizoma atractylodis macrocephalae and angelica;

(2) preparing silk fibroin;

(3) adding the extracts of the white atractylodes rhizome and the Chinese angelica into the silk fibroin solution to prepare the nano microspheres modified by the Chinese angelica and the white atractylodes rhizome.

2. A preparation method of nanometer microspheres of silk fibroin modified by angelica and bighead atractylodes rhizome is characterized in that:

s1, preparing the angelica sinensis extract: weighing radix Angelicae sinensis powder, adding methanol, performing ultrasonic treatment, adding methanol to complement weight, shaking, filtering, collecting filtrate, and drying to obtain radix Angelicae sinensis extract;

s2, preparing bighead atractylodes rhizome powder: adding 10 times of water into Atractylodis rhizoma, extracting at 100 deg.C for 2 times, mixing filtrates, concentrating under reduced pressure, vacuum drying, and pulverizing to obtain Atractylodis rhizoma powder;

s3, preparing the degumming silk: mixing Na ₂ CO ₃ Dissolving in 1L of ultrapure water, adding silk, stirring continuously at 100 ℃, taking out the silk after 30min, washing with ultrapure water to remove sericin and residual ions, repeating the operation for three times, loosening the silk cluster after degumming, and drying in a drying oven at 45-60 ℃ to obtain the degummed silk;

s4, silk fibroin: weighing the degumming silk, putting the degumming silk into a centrifugal tube, adding a lithium bromide solution, violently shaking, putting the solution into a water bath, heating to obtain a transparent viscous silk fibroin solution, centrifuging the obtained silk fibroin solution in a centrifugal machine for 1min, freezing, dialyzing, and freeze-drying;

3. The preparation method of the angelica and bighead atractylodes rhizome modified silk fibroin nanosphere as claimed in claim 2, which is characterized in that: removing insoluble substances and air bubbles in the solution in the step S4, filling into a dialysis bag, and dialyzing with ultrapure water for three days to completely remove Li in the solution ⁺ And Br ^— And pre-freezing the silk fibroin solution in a refrigerator at the temperature of-80 ℃ for 12 hours, and then putting the silk fibroin solution into a freeze dryer for freeze drying for 48 hours to obtain the silk fibroin fiber.

4. The preparation method of the angelica and bighead atractylodes rhizome modified silk fibroin nanosphere as claimed in claim 2, which is characterized in that: the concentration of the lithium bromide solution in the step S4 is 9.3 mol/L.

5. The preparation method of the angelica and bighead atractylodes rhizome modified silk fibroin nanosphere as claimed in claim 2, which is characterized in that: in the step S5, ethanol is quickly injected into a 3% silk fibroin solution according to the volume ratio of 1: 4, an ultrasonic cell disruptor is adopted to perform ultrasonic treatment under low power, the self-assembly is performed at room temperature, the solution is placed at-20 ℃ for completely freezing for 24 hours and then taken out, the solution is slowly melted into milky white solution at room temperature, and the emulsion is centrifuged at 4 ℃ and then is freeze-dried for later use.

6. The preparation method of the angelica and bighead atractylodes rhizome modified silk fibroin nanosphere as claimed in claim 2, which is characterized in that: in the step S5, the solution is incubated at 30 ℃ and 160r/min for 24 h.

7. The nano-microspheres prepared by the preparation method according to claims 2-6 can be applied to the field of cosmetics.