阅读说明：本技术 黄芪多糖在制备具有吸湿、保湿或舒缓功效的化妆品中的应用 (Application of astragalus polysaccharide in preparation of cosmetics with moisture absorption, moisture retention or relieving effects ) 是由 刘小花 王少华 刘珈铭 王昊 戴伟昊 于 2021-10-25 设计创作，主要内容包括：本发明提供黄芪多糖在制备具有吸湿、保湿或舒缓功效的化妆品中的应用。所述化妆品包括霜膏、乳液、膜粉剂、啫喱剂、粉饼、气雾剂和喷雾剂。所述黄芪多糖的制备方法为：将黄芪通过复合酶联合超声提取法进行提取,然后用乙醇沉淀,将乙醇回收后,再加入不同比例乙醇进行纯化,得到不同的黄芪多糖；所述复合酶为纤维素酶和木瓜蛋白酶。本发明的黄芪多糖具有良好的吸湿、保湿和舒缓功效,在化妆品领域具有一定的研究价值。应用本发明方法制备的多种醇沉黄芪多糖的吸湿、保湿和舒缓功效均佳。(The invention provides application of astragalus polysaccharide in preparing cosmetics with moisture absorption, moisture retention or relieving effects. The cosmetics comprise creams, lotions, film powders, gels, pressed powders, aerosols and sprays. The preparation method of the astragalus polysaccharide comprises the following steps: extracting radix astragali by complex enzyme combined ultrasonic extraction method, precipitating with ethanol, recovering ethanol, and purifying with ethanol of different proportions to obtain different astragalus polysaccharides; the complex enzyme is cellulase and papain. The astragalus polysaccharide disclosed by the invention has good moisture absorption, moisture retention and relieving effects, and has a certain research value in the field of cosmetics. The various alcohol-precipitated astragalus polysaccharides prepared by the method have good moisture absorption, moisture retention and relieving effects.)

1. Application of Astragalus polysaccharides in preparing cosmetic with moisture absorbing, moisturizing or relieving effects is provided.

2. Use according to claim 1, characterized in that: the cosmetics comprise creams, lotions, film powders, gels, pressed powders, mists, aerosols and sprays.

3. Use according to claim 1 or 2, characterized in that: the preparation method of the astragalus polysaccharide comprises the following steps: extracting radix astragali by complex enzyme combined ultrasonic extraction method, precipitating with above 90% ethanol, and recovering ethanol; adding ethanol for purification to obtain different ethanol-precipitated astragalus polysaccharides according to different ethanol contents in the solution; the complex enzyme is cellulase and papain.

4. Use according to claim 3, characterized in that: the cellulase is 2 percent, and the papain is 2 percent.

5. Use according to claim 3, characterized in that: the pH value of the complex enzyme extraction is 5.2, the temperature is 54.1 ℃, and the extraction time is 2 h.

6. Use according to claim 3, characterized in that: the ethanol precipitation is carried out by adding ethanol with the concentration of more than 90 percent until the ethanol concentration in the solution is 80 to 85 percent.

7. Use according to claim 3, characterized in that: the step of adding ethanol for purification is to add ethanol with the concentration of more than 90 percent, and then collect the precipitated astragalus polysaccharide when the concentration of the ethanol in the solution is 30 to 85 percent.

8. Use according to claim 7, characterized in that: the step of adding ethanol for purification is to add ethanol with the concentration of more than 90 percent, and then collect the precipitated astragalus polysaccharide when the concentration of the ethanol in the solution is 30 to 35 percent; or

The step of adding ethanol for purification is to add ethanol with the concentration of more than 90 percent, and then collect the precipitated astragalus polysaccharide when the concentration of the ethanol in the solution is 50 to 55 percent; or

The step of adding ethanol for purification is to add ethanol with the concentration of more than 90 percent, and then collect the precipitated astragalus polysaccharide when the ethanol concentration in the solution is 80 to 85 percent.

9. A cosmetic composition wherein the moisture-absorbing, moisturizing or soothing ingredient comprises the astragalus polysaccharides as described in claims 1 to 8.

10. The cosmetic according to claim 9, characterized in that: the radix hedysari polysaccharide with the effects of absorbing moisture, keeping moisture or relieving is prepared by adding ethanol into crude polysaccharide for purification, and collecting precipitated radix hedysari polysaccharide when the concentration of the ethanol in the solution is 30%, 50% or 80%.

Technical Field

The invention relates to application of astragalus polysaccharide in preparing cosmetics with moisture absorption, moisture retention or relieving effects.

Background

Polysaccharides are sugars in which 10 or more monosaccharides are linked by glycosidic bonds, and are also called polysaccharides. Polysaccharides are important forms of storage nutrients for plants, constitute the basic skeleton of plants, are important components of plants, and play important roles in plant bodies. Besides the action on the plant, the polysaccharide also has various unique biological activities of resisting tumor, inflammation and aging, treating cardiovascular and cerebrovascular diseases and the like. Plant polysaccharides are relatively less cytotoxic, and therefore plant polysaccharides are more and more appreciated and favored by people.

Radix astragali is the dry root of Astragalus membranaceus bge or Astragalus membranaceus bge of Leguminosae, and has effects of invigorating qi, consolidating superficial resistance, expelling toxin, expelling pus, promoting urination, and promoting granulation. Although astragalus and radix hedysari belong to the same family, the active ingredients of astragalus and radix hedysari are different after years of research. Astragalus polysaccharides are one of the main chemical components and main active components of Astragalus membranaceus. The dextran and heteropolysaccharide are main components of Astragalus polysaccharides, and have effects of regulating immunity, resisting oxidation and resisting aging.

With the increasing living standard, more and more people pay more attention to the self image and the prolonging of the service life in the modern life which is satisfied with the temperature saturation. With the increase of the demand of consumers for cosmetics, a large amount of cosmetics are introduced into the market, but some inferior cosmetics cause serious harm to consumers and society, and the case of disfiguration caused by chemical additives in some cosmetics causes consumers to talk about tiger color change. In order to ensure the safety of cosmetics, some synthetic chemicals are gradually replaced by natural products that are less cytotoxic. With the continuous development of technology and the continuous and deep research, more and more natural ingredients are added into cosmetics, so that the safety of the cosmetics is ensured, the curative effect of the cosmetics is improved, and consumers feel good at the cosmetics. The modern times and the science and technology are advanced, but the theme of the cosmetics is still bright, and the cosmetics with the effects of resisting aging, whitening, preventing sunlight and the like are durable.

Many natural products have been added to cosmetics to exert various effects. For example, the crude extract of paper mulberry fruit is separated by HPD-400 macroporous adsorption resin to obtain four parts of water, 30%, 60% and 95%, B16 cells are taken as research objects, how each part influences the activity of tyrosinase is researched, and the inhibition effect of each part on melanin synthesis is determined. The result shows that in the crude extract of the papermulberry fruit, 60 percent of ethanol part has the strongest effect of inhibiting the activity of tyrosinase, can inhibit the synthesis of melanin in B16 cells, and has better whitening effect. For another example, the method comprises the steps of extracting total flavonoids in the seabuckthorn seed meal by adopting microbial fermentation and secondary alcohol precipitation methods such as Shi right and the like, separating and purifying the extracted flavonoids after optimizing conditions of macroporous adsorption resin, and determining better process conditions by utilizing a single-factor test. The experimental result shows that the DPPH free radical clearance rate is in positive correlation with the content of the flavone in the seabuckthorn seed meal fermentation liquor, the seabuckthorn flavone basically has no toxic or side effect on fiber cells, the level of active oxygen can be obviously reduced, the content of matrix metalloproteinase is reduced, the content of collagen is increased, and the seabuckthorn seed meal fermentation liquor has a certain anti-aging effect.

When the water content in the horny layer of the skin is less than 10%, the skin becomes dry, the elasticity disappears, wrinkles appear, and the aging degree of the skin is remarkably accelerated. Therefore, the skin aging can be delayed by using the cosmetic with good moisture retention. And consumers are more receptive to the addition of natural products to cosmetics. At present, the research on astragalus polysaccharide mostly lies in the pharmacological action and mechanism in vivo, the pharmaceutical property and the application prospect in clinic, and the application of the astragalus polysaccharide in cosmetics is not seen yet.

Disclosure of Invention

In order to solve the problems in the prior art, the invention provides the application of astragalus polysaccharide in preparing cosmetics with the effects of absorbing moisture, preserving moisture or relieving. In the course of experiments, the subject group unexpectedly finds that astragalus polysaccharide is easy to absorb moisture and is not easy to store in daily environment, namely astragalus polysaccharide possibly has good moisture absorption. Therefore, the applicant researches the hygroscopicity and the moisture retention of astragalus polysaccharide through experiments, explores whether astragalus polysaccharide can replace the traditional cosmetic moisture retention agent, and hopes to develop a new direction for the research of traditional Chinese medicines.

In order to achieve the technical purpose of the invention, the invention provides the following technical scheme:

the invention provides application of astragalus polysaccharide in preparing cosmetics with moisture absorption, moisture retention or relieving effects.

Can be prepared into various cosmetics containing astragalus polysaccharide by the general method in the prior art, such as cream, emulsion, membrane powder, jelly, pressed powder, aqua, aerosol, spray and the like. The present invention is not limited to the preparation method of the above-mentioned various cosmetics.

Preferably, the preparation method of the astragalus polysaccharide comprises the following steps: extracting radix astragali by complex enzyme combined ultrasonic extraction method, precipitating with above 90% ethanol, and recovering ethanol; adding ethanol for purification to obtain different ethanol-precipitated astragalus polysaccharides according to different ethanol contents in the solution; the complex enzyme is cellulase and papain.

Preferably, the cellulase is 2% and the papain is 2%.

Preferably, the pH value of the complex enzyme extraction is 5.2, the temperature is 54.1 ℃, and the extraction time is 2 h.

Preferably, the ethanol precipitation is performed by adding ethanol with a concentration of 90% or more until the ethanol concentration in the solution is 80-85%.

Preferably, the ethanol is added for purification, wherein the ethanol with the concentration of more than 90 percent is added, and then the astragalus polysaccharide precipitated is collected when the ethanol concentration in the solution is 30 to 85 percent.

Preferably, the step of adding ethanol for purification is to add ethanol with the concentration of more than 90 percent, and then collect the precipitated astragalus polysaccharide when the concentration of the ethanol in the solution is 30 to 35 percent; or

The step of adding ethanol for purification is to add ethanol with the concentration of more than 90 percent, and then collect the precipitated astragalus polysaccharide when the concentration of the ethanol in the solution is 50 to 55 percent; or

The step of adding ethanol for purification is to add ethanol with the concentration of more than 90 percent, and then collect the precipitated astragalus polysaccharide when the ethanol concentration in the solution is 80 to 85 percent.

The invention also provides a cosmetic, wherein the component with the effects of absorbing moisture, keeping moisture or relieving comprises the astragalus polysaccharide.

Preferably, the hedysarum polybotrys polysaccharide with moisture absorption, moisture retention or soothing effects is hedysarum polybotrys polysaccharide which is precipitated when ethanol is added into crude polysaccharide for purification and the concentration of the ethanol in a solution is 30%, 50% or 80%.

The astragalus polysaccharide disclosed by the invention has good moisture absorption, moisture retention and relieving effects, and has a certain research value in the field of cosmetics. The 30%, 50% and 80% of three ethanol-precipitated astragalus polysaccharides prepared by the method have good moisture absorption, moisture retention and relieving effects.

Drawings

The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:

FIG. 1 is a standard curve of a glucose solution.

FIG. 2 shows the change of moisture absorption rate with time at RH of 43% (a) and 81% (b) for the Astragalus polysaccharides sample.

FIG. 3 shows the moisture retention rates of Astragalus polysaccharides and glycerol in different humidity environments.

FIG. 4 shows the results of the astragalan irritation test.

Detailed Description

The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from conventional biochemicals, unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.

The preparation method of the astragalus polysaccharide comprises the following steps:

weighing a proper amount of degreased radix astragali powder, adding cellulase (2 wt%, weight percentage of enzyme and acid water) and papain (2 wt%, weight percentage of enzyme and acid water), then adding acid water according to the water-material mass ratio of 30:1 (10, 10 and 10 times of solvent each time, extracting for three times), putting into a constant-temperature water bath pot, carrying out enzymolysis for 1.5-2.5 hours at 53.8-54.4 ℃, then carrying out ultrasonic extraction, filtering by using gauze, continuing to add acid water into the dregs for ultrasonic extraction for the second time and the third time, carrying out ultrasonic extraction for 30-40min each time, combining filtrates, centrifuging (8000 plus 10000r/min), taking supernatant, and concentrating to one tenth of the original volume. And (3) dropwise adding more than 90% of ethanol into the concentrated solution while stirring until the ethanol concentration is 80-85%. Standing, pouring out the filtrate, volatilizing the precipitate on a water bath until no ethanol smell exists, and freeze-drying to obtain the crude product of the astragalus polysaccharide.

When the concentration of ethanol in the solution is 80-85%, most of polysaccharide can be completely precipitated.

Taking a proper amount of the freeze-dried crude astragalus polysaccharide, adding a proper amount of water for dissolving, dropwise adding more than 90% of ethanol while stirring to enable the ethanol concentration to reach 30-35%, 50-55% and 80-85% respectively, centrifuging (8000r/min) to obtain precipitates, then obtaining 30-35%, 50-55% and 80-85% of astragalus alcohol precipitated polysaccharide, volatilizing the three parts of the alcohol precipitated polysaccharide on a water bath until no ethanol smell exists, freeze-drying, then placing in a dryer for storage, wherein the drying agent is allochroic silica gel.

The acid water is water adjusted to pH 5.2 with citric acid.

The above ethanol concentrations were measured at room temperature using an alcohol meter.

Example 1 study of moisture absorption and moisturizing efficacy of Astragalus polysaccharides

Test Material and Instrument

1.1 Experimental materials

Defatting radix astragali powder with 95% ethanol and ethyl acetate for 3 times to obtain defatted radix astragali powder.

1.2 enzymes and reagents for experiments

Table 1 shows the enzymes and reagents used in the extraction of Astragalus polysaccharides

1.3 Experimental instruments

TABLE 2 Experimental instruments for extracting Astragalus polysaccharides

2 method of experiment

2.1 preparation of Astragalus polysaccharides

Weighing 3200g of defatted astragalus powder, adding 64g of cellulase and 64g of papain, adding 32000g of acid water, putting into a constant-temperature water bath, carrying out enzymolysis for 2 hours at 54.1 ℃, then carrying out ultrasonic extraction, filtering by using gauze after extraction, continuously adding acid water into medicine dregs, carrying out ultrasonic extraction for the second time and the third time, carrying out ultrasonic extraction for 30min each time, adding 32000g of acid water into medicine dregs each time, combining filtrates, centrifuging (8000r/min), taking supernatant, and concentrating to one tenth of the original volume. And (3) dropwise adding 90% ethanol into the concentrated solution while stirring until the ethanol concentration is 80%. Standing, pouring out the filtrate, volatilizing the precipitate on a water bath until no ethanol smell exists, and freeze-drying to obtain the crude product of the astragalus polysaccharide.

Taking a proper amount of the freeze-dried crude astragalus polysaccharide, adding a proper amount of water for dissolving, dropwise adding 90% ethanol while stirring to enable the ethanol concentration to reach 30%, 50% and 80% respectively, centrifuging (8000r/min) to obtain precipitates, then obtaining 30%, 50% and 80% of astragalus ethanol precipitated polysaccharide, volatilizing the three parts of the ethanol precipitated polysaccharide on a water bath until no ethanol smell exists, freeze-drying, then placing into a dryer for storage, wherein the drying agent is allochroic silica gel.

The acid water is water adjusted to pH 5.2 with citric acid.

The above ethanol concentrations were measured at room temperature using an alcohol meter.

2.2 determination of Astragalus polysaccharides content

2.2.1 preparation of glucose Standard Curve and results

Weighing 25mg of glucose, adding water to prepare 100mL of glucose standard solution, precisely sucking 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL and 1.2mL of glucose standard solution, placing the glucose standard solution in a 10mL volumetric flask, sucking 2mL of the glucose standard solution respectively after constant volume, placing the glucose standard solution in a test tube, adding 1mL of newly-prepared 5% phenol solution, mixing uniformly, adding 5mL of 98% concentrated sulfuric acid, vortexing, and cooling to room temperature by tap water after 15 min. The absorbance of each solution after the reaction was measured at 490nm with an ultraviolet spectrophotometer using distilled water as a blank control in the above manner. And (3) taking the concentration of the glucose solution as an abscissa and the absorbance of the solution as an ordinate to prepare a standard curve, and calculating to obtain a regression equation.

The regression equation of the glucose standard curve is as follows: 16.722x-0.0146 (R)²＝0.992)。

FIG. 1 is a standard curve of a glucose solution.

TABLE 3 glucose Standard Curve for the determination of results

2.2.2 determination of polysaccharide content and yield

Weighing 25mg of each polysaccharide sample, dissolving the polysaccharide sample with a proper amount of distilled water, placing the dissolved polysaccharide sample into a 100mL volumetric flask, and fixing the volume. And (3) measuring the absorbance of the sample solution according to the preparation method of the standard curve in the step 2.2.1, and substituting the absorbance into the standard curve to calculate the concentration of the polysaccharide in the sample solution. And calculating the mass of the polysaccharide in the sample according to the concentration of the polysaccharide in the sample solution, wherein the ratio of the mass of the polysaccharide to the mass of the weighed sample is the content of the polysaccharide in the sample.

2.3 moisture absorption test

Putting a weighing bottle in an oven for 6h, accurately weighing 0.5g of freeze-dried 30%, 50% and 80% of astragalus membranaceus alcohol precipitation polysaccharide sample prepared in the step 2.1 and 3 parts of reference glycerol, respectively adding the samples and the reference glycerol into the weighing bottle after the constant weight, setting a temperature of a drug stability test box to be 20 ℃, and Relative Humidity (RH) to be 43%, putting the drug stability test box into the weighing bottle containing the samples after the drug stability test box is stable, placing the bottles for 1h, 3h, 6h, 10h, 24h, 34h and 48h, accurately weighing the mass of each weighing bottle, and calculating the moisture absorption rate according to the mass difference of each sample and glycerol before and after moisture absorption. Moisture absorption rate ═ M_n-M₀)/M₀×100％，M₀For the mass of each sample before moisture absorption, M_nFor each sample in a drug stability test chamberThe mass after the corresponding time.

And continuously placing each sample in a high-humidity drug stability test box with the temperature of 20 ℃ and the RH of 43 percent until the moisture absorption of each sample reaches saturation.

And accurately weighing 3 parts of dried 30%, 50% and 80% astragalus polysaccharide samples and reference glycerol, respectively adding the weighed samples into weighing bottles with constant weight, setting the temperature of a drug stability test box to be 20 ℃, setting RH to be 81%, and measuring the moisture absorption rate of each sample when the RH is 81% by the same measuring step of the moisture absorption rate when the RH is 43%.

Each sample was kept in a high humidity drug stability test chamber at 20 ℃ and an RH of 81% until each sample was saturated by moisture absorption.

2.4 moisture Retention test

And (3) continuously placing the samples with the RH of 43% and the RH of 81% respectively reaching moisture absorption saturation in the step 2.3 in a medicine stability test box with the temperature of 20 ℃ and the RH of 43% and 81% respectively, placing for 48h, accurately weighing the mass of each sample, and calculating the moisture retention rate of each sample according to the moisture absorption mass difference before and after the samples are placed. Moisture retention rate ═ C_n/C₀X 100%, wherein C₀The amount of moisture absorption after saturation of moisture absorption of each sample, C_nThe amount of moisture absorbed by each sample after standing for 48 hours.

3 results and discussion

3.1 Astragalus polysaccharides content and yield

The polysaccharide yields of the various fractions of Astragalus are shown in Table 4, and the polysaccharide contents and yields are shown in Table 5. From table 5, it can be seen that the highest content of astragalus polysaccharides is 80% alcohol precipitated polysaccharides, and the lowest content is 30% alcohol precipitated polysaccharides; the highest yield is 80% of alcohol precipitated polysaccharide, and the lowest yield is 50% of alcohol precipitated polysaccharide.

The polysaccharide content is CV/1000M × 100%, and C in the formula is the mass concentration (mg.l-1) of the polysaccharide solution obtained by the regression equation; v is the volume (mL) of the freeze-dried polysaccharide sample with constant volume; sample amount of lyophilized polysaccharide (mg).

The yield of polysaccharide was M/1000M × 100%, where M is the mass (mg) of the lyophilized polysaccharide sample; m is the mass (g) of the defatted astragalus powder.

TABLE 4 yield of each product in the preparation of Astragalus polysaccharides

TABLE 5 content and yield of polysaccharides from various parts of Astragalus membranaceus

3.2 hygroscopicity of Astragalus polysaccharides

The change in the moisture absorption of glycerin with time at RH of 43% and 81% is shown in table 6.

TABLE 6 relationship between the amount of glycerin absorbed moisture in an environment with 43% RH as a function of time

3.2.1 hygroscopicity of Astragalus polysaccharides

The changes in moisture absorption with time of each fraction of astragalus polysaccharides at RH of 43% and 81% are shown in tables 7 and 8.

TABLE 7 relationship between moisture absorption of Astragalus polysaccharides in each part of Astragalus polysaccharides with time in 43% RH environment

TABLE 8 relationship between moisture absorption of Astragalus polysaccharides in each part of Astragalus polysaccharides with time in an environment with RH of 81%

The moisture absorption rate of astragalus membranaceus in 30% alcohol precipitated polysaccharide, 50% alcohol precipitated polysaccharide, 80% alcohol precipitated polysaccharide and glycerol in 43% and 81% RH environments along with the change of time curves are shown in fig. 2.

FIG. 2 shows the change of moisture absorption rate with time at RH of 43% (a) and 81% (b) for the Astragalus polysaccharides sample.

As can be seen from the a diagram of fig. 2, the moisture absorption rate of the three astragalus polysaccharides is significantly lower than that of glycerin when the ambient humidity is relatively low (RH ═ 43%). The moisture absorption rates of the first three astragalus polysaccharides are in the order of 30% alcohol precipitated polysaccharide to 80% alcohol precipitated polysaccharide to 50% alcohol precipitated polysaccharide, the moisture absorption rates of the last three astragalus polysaccharides are in the order of 50% alcohol precipitated polysaccharide to 30% alcohol precipitated polysaccharide to 80% alcohol precipitated polysaccharide, and the difference of the moisture absorption rates of the first three astragalus polysaccharides is larger than that of the last three astragalus polysaccharides after 24 h. The moisture absorption rates of 30% of alcohol precipitation polysaccharide and 80% of alcohol precipitation polysaccharide increase with time before 10 hours, the maximum moisture absorption rate reaches 10 hours, the maximum moisture absorption rate of 30% of alcohol precipitation polysaccharide is 5.98%, and the maximum moisture absorption rate of 80% of alcohol precipitation polysaccharide is 5.61%. The moisture absorption rates of these two astragalus polysaccharides subsequently decreased and then increased with increasing time. The moisture absorption rate of the 50% alcohol-precipitated polysaccharide in the determination time range is increased along with the increase of time and reaches a maximum value within 48 hours, and the maximum moisture absorption rate of the 50% alcohol-precipitated polysaccharide is 5.90%. The three astragalus polysaccharides basically reach moisture absorption saturation after 48 hours.

As can be seen from the b-diagram of fig. 2, at relatively high ambient humidity (RH 81%), all three astragalus polysaccharides have a certain hygroscopicity, but still significantly less than the moisture absorption rate of glycerin. The moisture absorption rates of the first three astragalus polysaccharides are in the order of 30% alcohol precipitated polysaccharide > 80% alcohol precipitated polysaccharide > 50% alcohol precipitated polysaccharide, and the moisture absorption rates of the last three astragalus polysaccharides are in the order of 80% alcohol precipitated polysaccharide > 30% alcohol precipitated polysaccharide > 50% alcohol precipitated polysaccharide. The moisture absorption rates of the three astragalus polysaccharides before 24 hours are increased continuously along with the increase of time, the moisture absorption rates of the three astragalus polysaccharides after 24 hours reach the maximum value, at the moment, the maximum moisture absorption rate of 30% alcohol precipitation polysaccharide is 20.24%, the maximum moisture absorption rate of 50% alcohol precipitation polysaccharide is 19.26%, and the maximum moisture absorption rate of 80% alcohol precipitation polysaccharide is 23.43%. After 24h, the moisture absorption rates of the three astragalus polysaccharides are reduced and then increased along with the increase of time, and after 48h, the three astragalus polysaccharides basically reach a saturated state.

It can be seen from fig. 2 that the hygroscopicity of the 30% alcohol precipitated polysaccharide sample is relatively better in the low humidity environment, and the hygroscopicity of the 80% alcohol precipitated polysaccharide sample is better than that of the other two polysaccharide samples in the high humidity environment.

3.2.2 analysis of variance of moisture absorption rates of Astragalus polysaccharides and Glycerol

The results of the analysis of variance of the moisture absorption rates of astragalan and the control glycerin at RH of 43% and 81% are shown in table 9. As can be seen from the results in the table, the difference in moisture absorption rates between astragalus polysaccharide and glycerin was statistically significant at an RH of 43% and 81% as α of 0.05, and it was considered that the total average moisture absorption rates of astragalus polysaccharide and glycerin were not all equal. As can be seen from fig. 2, glycerin has the highest moisture absorption rate.

TABLE 9 analysis of moisture absorption rate variance of Astragalus polysaccharides and Glycerol in different RH environments

3.3 moisturizing Properties of Astragalus polysaccharides

3.3.1 moisturizing Properties of Astragalus polysaccharides

The moisturizing rates of 30% alcohol precipitated polysaccharide, 50% alcohol precipitated polysaccharide, 80% astragalus polysaccharide and glycerin of astragalus in different humidity environments are shown in fig. 3 and table 10. From the data in table 10 and fig. 3, it can be seen that in the relative humidity environment of 43%, the moisture retention rates of the three astragalus polysaccharides and glycerin are in the order of 30% alcohol precipitated polysaccharide > 80% alcohol precipitated polysaccharide > glycerin > 50% alcohol precipitated polysaccharide. In an environment with 81% of relative humidity, the moisture retention rate of 80% of alcohol-precipitated polysaccharide is the largest and reaches 98.21%, the moisture retention rate of 50% of astragalus membranaceus is the worst, and the moisture retention rate of the three astragalus membranaceus polysaccharides is slightly smaller than that of glycerin.

FIG. 3 shows the moisture retention rates of Astragalus polysaccharides and glycerol in different humidity environments.

TABLE 10 moisturizing rates of Astragalus polysaccharides and Glycerol in different humidity environments

3.3.2 analysis of variance of moisture absorption rates of Astragalus polysaccharides and Glycerol

The results of the anova analysis of the moisturizing ratio of astragalan and the control glycerin at RH of 43% and 81% are shown in table 11. From the results, it was found that the difference in moisture absorption rates between astragalus polysaccharide and glycerin was statistically significant at an RH of 43% at a level of α of 0.05, and the total average moisture retention rates of astragalus polysaccharide and glycerin were not all equal.

TABLE 11 moisture retention analysis of variance of astragalus polysaccharides and glycerol in different RH environments

3.4 final phrase

The highest yield and highest content of astragalus polysaccharides are all 80% of alcohol precipitated polysaccharides. The moisture absorption experiment result shows that when the RH is 43% at 20 ℃, the maximum moisture absorption rate of the astragalus polysaccharide is 30% of the ethanol precipitated polysaccharide part, and is 5.98%; when the RH is 81%, the moisture absorption rate of 80% alcohol precipitation polysaccharide in the astragalus polysaccharide is the maximum, and reaches 23.43%. At 20 ℃ and 43% RH, the ethanol precipitated polysaccharide fraction with the maximum moisture retention rate of 30% in the astragalus polysaccharides is 36.76%; when the RH is 81%, the moisture retention rate of 80% of alcohol precipitation polysaccharide in the astragalus polysaccharide is the maximum, and reaches 98.21%.

The results of the anova showed that the hygroscopicity of astragalus polysaccharides was statistically different from that of glycerol at RH of 43% and 81%, and the hygroscopicity of astragalus polysaccharides was different from that of glycerol. When the RH is 81%, the astragalus polysaccharide has good moisture retention.

Astragalus membranaceus is widely applied as a traditional Chinese medicinal material in China and a good medicine for tonifying qi, and the pharmacological action and the action mechanism of astragalus polysaccharide are always hot spots of research. The application develops a new method, researches the moisture absorption and moisture preservation effects of the astragalus polysaccharide, provides a theoretical basis for further applying the astragalus polysaccharide to cosmetics, and develops new application of the traditional Chinese medicine.

Example 2 relief efficacy of Astragalus polysaccharides component (acute irritation test results for Zebra fish)

On the basis of the evaluation of the early stage moisturizing property, the applicant continuously refers to the standard of 'cosmetic safety experiment (zebra fish) experimental method' issued by the bodhumva cosmetic industry association in the Guangzhou development area, and the soothing effect of the related traditional Chinese medicine polysaccharide components is evaluated through a mild irritation test.

The experimental method comprises the following steps: zebra fish embryo determination method for mild irritation of cosmetics

1 this standard specifies the methodology and operation of the zebra fish embryo experiment for evaluating the mild irritation of cosmetics. The standard is suitable for mild irritation evaluation of cosmetics (cream, emulsion, gel, essence, etc.) and cosmetic raw materials (liquid water base, cream emulsion, gel, soluble powder).

2 cosmetic safety specifications (2015 edition) QB/T1684-2015 cosmetic inspection rules DB32/T3979-2021 Zebra fish feeding technical conditions OECD236 Fisherbiroaceuteoxicity (FET) Test for experiments.

Principle of 3 method

24hpf zebrafish embryos are placed in different samples under blank control or positive control conditions by using a 24-well cell culture plate, observed under an integral microscope within 5min, and the number of times of stimulation swing (movement of yolk sac position) and the death (coagulation) amount of the fish embryos are recorded according to the mild stimulation response of the embryos to the tested samples (the mild stimulation response comprises stimulation swing and coagulation (death) of the embryos) so as to characterize the mild stimulation of the samples.

4 biological model

4.1 adult fish spawns by adopting wild zebrafish (Daniorio) adult fish (4-18 months) which is sexually mature, healthy, free from deformity, high in spawning amount and good in spawning quality. Adult fish has a body length of 3-5cm, and male fish has a slender body shape, is lemon-colored, and has a flat abdomen. The female fish is full of body, the abdomen is enlarged and silvery, and the body color is silvery gray.

4.2 the fish embryo adopts fertilized eggs which normally develop at the 24hpf period, and the object identification refers to the appendix A of the method for measuring the mild irritation of cosmetics and the zebra fish embryo.

4.3 culture and maintenance of fish and maintenance reference < appendix F of the method for measuring the mild irritation of cosmetics and the embryo of zebra fish >.

5 reagent of material

5.1 materials

5.1.1 Zebra fish mating box. An embryo collector: 100.0mm petri dish. 24-well cell culture plate: the volume of each hole is 3.5 ml. Plastic dropper: 3.0 ml.

5.1.2 analytically pure reagents: sodium chloride (NaCl), potassium chloride (KCl), calcium chloride (CaCl)₂·2H₂O), magnesium sulfate (MgSO)₄·7H₂O)。

5.2 control group solution

5.2.1 blank control group: standard dilution water. The preparation method of the standard dilution water comprises the following steps: precisely weighing 17.2g of sodium chloride, 0.76g of potassium chloride, 4.9g of magnesium sulfate and 2.9g of calcium chloride, dissolving the sodium chloride, the potassium chloride, the magnesium sulfate and the calcium chloride by using distilled water or deionized water, and fixing the volume to 1000ml to obtain a standard dilution water stock solution. Taking 16.67ml of the standard dilution water stock solution, and then diluting the stock solution to 1000ml by using distilled water or deionized water.

5.2.2 Positive control group: sodium Dodecyl Sulfate (SDS) standard stock solution (5.0 mg/ml).

6 Experimental procedures

6.1 embryo preparation

6.1.1 when the zebra fish is bred, 2/3 volumes of culture water are added in advance in an outer tank of an egg laying box, then an inner tank with embryo separation function is sleeved, a baffle is inserted, and male and female fishes are separated. Selecting sexual maturity zebra fish to be bred for more than 4 months in the evening before the experiment, and placing the zebra fish in a spawning box with a baffle in a light-proof overnight according to a male-female ratio of 1:1 or 1: 2.

6.1.2 the next morning, the light source is turned on and then the baffle of the spawning box is pulled out to mate and spawn for 1 h. Then checking the spawning condition of each jar of adult fish, collecting embryos by using a strainer spoon, washing the embryos by using breeding water so as to place the embryos in culture dishes, wherein the number of the embryos in each culture dish is not more than 300 on average, and then placing the culture dishes into a constant-temperature incubator for incubation.

6.2 pretreatment of samples refer to zebra fish embryo method annex D cosmetics and raw material pretreatment methods.

6.3 Experimental methods

6.3.1 Experimental procedures embryos with normal 24hpf development were randomly picked with a plastic dropper and placed under a stereomicroscope for testing (at this time, embryos at somite stage). The method specifically comprises the following steps: adding 10-20 embryos into each hole of the cell culture plate, sucking culture water with a plastic straw, adding 1.0ml of prepared different sample solutions into each hole, and standing in the 24-hole cell culture plate. Each concentration group was provided with 3 parallel groups, and a blank control group or a positive control group was provided. The embryo stimulated swing (movement of yolk sac position) and coagulation (death) were observed and recorded under a stereomicroscope (objective 2-3 times) within 5 min.

6.3.2 judging standard test end point and evaluation index by taking embryo coagulation and 5min as test end points, the expression form is as follows: embryos stimulate either conditions of oscillation, coagulation (death).

6.3.3 the result requires that the fish embryo should be positioned in the middle of the video picture, so that the fish embryo is not damaged, and the fish embryo can normally carry out formal experiments.

7 result of calculation

7.1 record and calculate the cumulative number of stimulation swings and the cumulative mortality rate of embryos (24hpf) of each parallel group in each group within 5 min. Wherein, the cumulative mortality rate is 100% of the cumulative number of dead embryos/total number of embryos.

7.2 statistical analysis of cumulative stimulation swing times and cumulative mortality was performed using statistical analysis software (such as GraphPadprism8), values expressed as Mean. + -. standard error (Mean. + -. SEM).

8 quality control

The test result meets the following requirements, and the result is effective. Otherwise, the experiment should be repeated after the cause is found:

8.1 within 5min of blank control test, the death rate of the zebra fish embryo of the blank control group is less than or equal to 10 percent.

8.2 the embryo death rate of zebra fish in the positive control group is more than 90% within 5min of the positive control test.

9 waste disposal test zebra fish embryos are treated as general waste after being inactivated in ice water at 2-8 ℃. Samples with mild irritancy, waste solutions containing Sodium Dodecyl Sulfate (SDS) were disposed of as hazardous waste.

(II) results of the experiment

In the irritation test, the astragalus component polysaccharides showed no irritation to zebrafish (table 12 and figure 4).

Fig. 4 shows the results of the cistanche polysaccharide irritation test (the values in fig. 4 are the average values of three groups).

In the mild irritation experiments, each sugar of astragalus polysaccharides was not irritating by anova (table 13).

TABLE 12 results of irritation test (unit: times/min)

TABLE 13 irritation test ANOVA results

Summary of the invention

The experimental results are combined with the previous moisturizing effect experiments, and the results show that the 30%, 50% and 80% ethanol precipitation components of the astragalus polysaccharide have certain effects on moisturizing, no obvious stimulation effect exists in the main components, and the astragalus polysaccharide has a value of continuing further research by combining the effects of the astragalus traditional Chinese medicinal materials on health care.

EXAMPLE 3 preparation of Astragalus polysaccharides

Weighing an appropriate amount of 1000g of defatted astragalus powder, adding 20g of cellulase and 20g of papain, then adding 10000g of acid water, putting the mixture into a constant-temperature water bath, carrying out enzymolysis for 1.5 hours at 54.4 ℃, then carrying out ultrasonic extraction, filtering by using gauze, continuously adding acid water into medicine dregs, carrying out ultrasonic extraction for the second time and the third time, carrying out ultrasonic extraction for 40min each time, adding 10000g of acid water each time, combining filtrates, then centrifuging (9000r/min), taking supernatant, and concentrating to one tenth of the original volume. 95% ethanol is added dropwise to the concentrated solution while stirring, and finally the ethanol concentration is 85%. Standing, pouring out the filtrate, volatilizing the precipitate on a water bath until no ethanol smell exists, and freeze-drying to obtain the crude product of the astragalus polysaccharide.

Taking a proper amount of the freeze-dried crude astragalus polysaccharide, adding a proper amount of water for dissolving, dropwise adding 95% ethanol while stirring to enable the ethanol concentration to reach 35%, 55% and 85% respectively, centrifuging (9000r/min) to obtain precipitates, then obtaining 35%, 55% and 85% of astragalus ethanol precipitated polysaccharide, volatilizing the three parts of the ethanol precipitated polysaccharide on a water bath until no ethanol smell exists, freeze-drying, then placing into a dryer for storage, wherein the drying agent is allochroic silica gel.

The acid water is water adjusted to pH 5.2 with citric acid.

The above ethanol concentrations were measured at room temperature using an alcohol meter.

Example 4 preparation of Astragalus polysaccharides

Weighing an appropriate amount of 1000g of defatted astragalus powder, adding 20g of cellulase and 20g of papain, then adding 10000g of acid water, putting the mixture into a constant-temperature water bath, carrying out enzymolysis for 2.5 hours at 53.8 ℃, then carrying out ultrasonic extraction, filtering by using gauze, continuously adding acid water into medicine dregs, carrying out ultrasonic extraction for the second time and the third time, carrying out ultrasonic extraction for 35min each time, adding 10000g of acid water into the medicine dregs, combining the filtrate, then centrifuging (10000r/min), taking supernatant, and concentrating to one tenth of the original volume. And (3) dropwise adding 98% ethanol into the concentrated solution while stirring until the ethanol concentration is 82%. Standing, pouring out the filtrate, volatilizing the precipitate on a water bath until no ethanol smell exists, and freeze-drying to obtain the crude product of the astragalus polysaccharide.

Taking a proper amount of the freeze-dried crude astragalus polysaccharide, adding a proper amount of water for dissolving, dropwise adding 95% ethanol while stirring to enable the ethanol concentration to reach 30%, 55% and 80% respectively, centrifuging (10000r/min) to obtain precipitates, then obtaining 30%, 55% and 80% of astragalus ethanol precipitated polysaccharide, volatilizing the three parts of the ethanol precipitated polysaccharide on a water bath until no ethanol smell exists, freeze-drying, then placing into a dryer for storage, wherein the drying agent is allochroic silica gel.

The acid water is water adjusted to pH 5.2 with citric acid.

The above ethanol concentrations were measured at room temperature using an alcohol meter.

Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.