CN112972321B - Natural antioxidant for cosmetics and preparation method thereof - Google Patents
Natural antioxidant for cosmetics and preparation method thereof Download PDFInfo
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- CN112972321B CN112972321B CN201911296785.0A CN201911296785A CN112972321B CN 112972321 B CN112972321 B CN 112972321B CN 201911296785 A CN201911296785 A CN 201911296785A CN 112972321 B CN112972321 B CN 112972321B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/52—Stabilizers
- A61K2800/522—Antioxidants; Radical scavengers
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/82—Preparation or application process involves sonication or ultrasonication
Abstract
The invention discloses a natural antioxidant for cosmetics and a preparation method thereof. Different components have different characteristics in activity, water solubility and cost accounting, the factors of activity, solubility, cost and the like are combined, a compounding scheme of various components can be adopted, and particularly, the mass ratio of polysaccharide to isoflavone is 1:2, the synergistic effect between the two components is the best, and the activity is the highest. The red clover is widely distributed in the world as leguminous forage, has rich sources, and has health care function of isoflavone components. The product is easy to prepare, safe and environment-friendly in process and low in cost, and the formula is diversified, so that the product can meet the requirements of different purposes, dosage forms and the like, and has a wide application range in cosmetics as a plant source antioxidant.
Description
Technical Field
The invention relates to the technical field of cosmetic industry, in particular to a natural antioxidant for cosmetics and a preparation method thereof.
Background
The skin is the outermost tissue organ of the human body, plays a role in protecting the human body from external irritation or injury, and is the tissue which can reflect the aging of the organism at the earliest time. The factors causing skin aging are many, except genetic factors, the skin aging comes from external environments such as ultraviolet rays and haze, and in addition, mental pressure and irregular daily work and rest inside a body can generate a large amount of free radicals to interfere cell signal conduction, accelerate apoptosis and accelerate skin aging, and the skin aging is particularly characterized by dry, loose, scurf, increased melanin and even colored spots and the like of the skin, and even causes skin cancer, thereby affecting the health and life quality of people.
The antioxidant serving as an exogenous antioxidant can effectively eliminate oxygen free radicals, so that the skin achieves the purposes of ageing resistance, whitening, sun protection, moisture preservation and the like. Along with the continuous improvement of the national quality of life, people pay more and more attention to the maintenance of the body, cosmetics are not just patents of young women any more, the demand is large, and the requirements on products tend to be diversified.
Antioxidants are substances that at certain concentrations prevent or inhibit oxidative damage by scavenging free radicals. At present, tert-butyl p-hydroxyanisole (BHA) and dibutyl hydroxy toluene (BHT) are widely used in synthetic antioxidants. However, in the experiment of intraperitoneal injection of BHA, BHT and its metabolites into young male mice, the number of mouse lung cells was found to be increased, and the use of such synthetic antioxidants has been gradually prohibited in the United states, countries of the European Union, etc.
The substitution of natural antioxidants for synthetic antioxidants is a trend in the future. Cosmetics are substances that are in continuous contact with human skin for a long time, and must be mild, safe, non-toxic, and free of side effects. Therefore, the natural antioxidant with high efficiency, low toxicity and low price is indispensable for safe and healthy cosmetics.
Disclosure of Invention
The invention aims to provide a natural antioxidant for cosmetics and a preparation method thereof, and the antioxidant component extracted and prepared from the plant has better activity than an isoflavone component, better water solubility, wider application range and very good development and application prospects, so as to solve the problems in the background technology.
In order to achieve the purpose, the invention provides the following technical scheme: a natural antioxidant for cosmetic comprises extract of Trifolium pratense extract, and contains crude polysaccharide, crude isoflavone, refined polysaccharide, refined isoflavone and compound of refined polysaccharide and refined isoflavone.
Further, the mass ratio of the compound of the refined polysaccharide and the refined isoflavone is 1:10-15:1, preparation.
The invention provides another technical scheme: a method for preparing natural antioxidant for cosmetic comprises the following steps:
s1: preparation of red clover extract
Deactivating enzyme of collected red clover leaves at 105 deg.C for 2-60min, drying at 65 deg.C for 1-160h, pulverizing, defatting with small amount of petroleum ether for 1-10 times, volatilizing solvent, air drying, weighing certain mass of defatted red clover powder, ultrasonic extracting with 10-50 times of 10-100% ethanol at 30-70 deg.C for 10-90min, filtering, collecting supernatant, extracting sample residue with 5-120 times of solvent for 10-90min, and mixing the two extractive solutions;
s2: preparation of crude polysaccharide and crude isoflavone
Crude polysaccharide: concentrating the extractive solution at 15-90 deg.C with vacuum rotary instrument until the volume is one eighth, adding 2-20 times volume of anhydrous ethanol into the concentrated solution under stirring, standing in 4 deg.C refrigerator overnight, centrifuging at 4800rpm for 2-20min, centrifuging to obtain brown precipitate as crude polysaccharide part, repeatedly washing the precipitate with anhydrous ethanol for 1-10 times, washing to remove alcohol soluble impurities, washing the precipitate with ethanol, pouring into culture dish, and volatilizing solvent to obtain brown red clover crude polysaccharide;
the supernatant is the isoflavone part, and the supernatant is decompressed and concentrated to obtain crude isoflavone of the red clover;
s3: refining of red clover isoflavone
Soaking the pretreated resin in distilled water for more than 10-90h, loading the column resin in wet method at a height of 3-50cm, dissolving 1-30g of crude isoflavone in water, loading the sample, waiting for adsorption for 0.5-24h, eluting with distilled water, 30% and 50% ethanol for 1-5BV respectively, eluting with 70% ethanol for 1-5BV, finally eluting with 100% ethanol for 1-10BV, collecting 70% ethanol eluate, and concentrating under reduced pressure to obtain refined isoflavone;
s4: refining of red clover polysaccharide
Protein removal: preparing 1-10mg/mL solution of red clover crude polysaccharide with distilled water, adding 1/4 volume of sevage solution, oscillating for 1-30min, transferring to a separating funnel, standing for 1-120min, discharging the lower layer solution and the middle layer protein, and repeating the operation for 1-6 times until the protein does not appear in the lower layer organic solvent;
and (3) dialysis: concentrating the protein-removed polysaccharide solution at low temperature under reduced pressure, filling into a pretreated dialysis bag, placing into a large-capacity beaker, filling distilled water, standing for dialysis, changing water every 1-16h, dialyzing for 1-72h, and checking residual conditions of NaCl, KCl and the like by using 1% AgNO3 to finally obtain the purified polysaccharide.
Further, for step S3, resin pretreatment: soaking the resin in 20-100% ethanol for 1-72h, filtering, repeatedly washing with deionized water until there is no alcohol smell and the washing solution is white and turbid, soaking in 1-30% NaOH for 1-72h, filtering, repeatedly washing the resin with deionized water until the pH value is neutral, soaking in 1-30% HCl solution for 1-72h, filtering and washing the resin to neutral, and drying at 20-90 deg.C for use.
Compared with the prior art, the invention has the beneficial effects that:
the antioxidant component extracted and prepared from the plant has high activity, good water solubility, wide application range, greenness, safety and health care function. In addition, the red clover plant has rich resources, low development cost and good market prospect.
Detailed Description
The following examples will explain the present invention in detail, however, the present invention is not limited thereto. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
A natural antioxidant for cosmetic comprises crude Trifolium pratense polysaccharide and crude isoflavone, wherein:
the raw material proportion of the red clover crude polysaccharide and the crude isoflavone is as follows: 50g of red clover leaves, 0.2g of degreased petroleum ether, 5g of 40% ethanol and 4g of absolute ethanol.
Based on the above natural antioxidant for cosmetics, this example proposes a method for preparing a natural antioxidant for cosmetics, comprising the steps of:
s101: deactivating enzyme at 105 deg.C for 10min, drying at 65 deg.C for 72 hr, pulverizing, defatting with small amount of petroleum ether for 4 times, volatilizing solvent, and air drying;
s102: weighing 2g of defatted red clover powder, performing ultrasonic assisted extraction with 25 times of 40% ethanol at 50 ℃ for 30min, filtering and collecting supernatant;
s103: adding 20 times of solvent into the sample residue, extracting for 30min, and mixing the two extractive solutions;
s104: concentrating the extractive solution at 45 deg.C with vacuum rotary instrument until the volume is one eighth, slowly adding 4 times of anhydrous ethanol into the concentrated solution under stirring to make the ethanol concentration reach 80%, placing in 4 deg.C refrigerator, standing overnight, centrifuging at 4800rpm for 10min, centrifuging to obtain brown precipitate as crude polysaccharide part and supernatant as isoflavone part;
s105: washing the precipitate with anhydrous ethanol for 3 times, washing off alcohol soluble impurities, washing out the precipitate with ethanol, pouring into a culture dish, volatilizing the solvent to obtain brown crude Trifolium pratense polysaccharide with extraction rate of 2.26%, and concentrating the supernatant under reduced pressure to obtain crude Trifolium pratense isoflavone 320.62mg with extraction rate of 16.3%.
Example 2
The red clover crude isoflavone sample contains monosaccharide, oligosaccharide and inorganic salt, and needs to be further refined.
The refining of the red clover crude isoflavone comprises the following steps:
pretreatment of AB-8 macroporous adsorption resin: soaking the resin in 95% ethanol for 24h, filtering, repeatedly washing with deionized water until no alcohol smell exists, and the washing liquid has no white turbidity, soaking in 5% NaOH for 24h, filtering, repeatedly washing the resin with deionized water until the pH value is neutral, soaking in 5% HCl solution for 24h, filtering and washing the resin to neutral, and drying at 60 ℃ for later use;
refining: soaking the pretreated resin in distilled water for more than 48h, packing with wet method to obtain column with resin height of 13cm, dissolving crude isoflavone 10g in water, loading sample, waiting for adsorption for 2h, eluting with distilled water, 30% and 50% ethanol respectively for 1BV, eluting with 70% ethanol for 2BV, eluting with 100% ethanol for 1BV, collecting 70% ethanol eluate, and concentrating under reduced pressure to obtain refined isoflavone 1.73g with isoflavone content of 42%.
Example 3
The red clover crude polysaccharide sample contains protein, monosaccharide, oligosaccharide and inorganic salt, and needs to be further refined.
The refining of the red clover crude polysaccharide comprises the following steps:
protein removal: preparing 2mg/mL solution of red clover crude polysaccharide with distilled water, adding 1/4 volume of sevage solution (n-butanol: chloroform 1:4), oscillating for 20min, transferring to a separating funnel, standing for 20min, discharging the protein in the lower layer solution and the middle layer, and repeating the operation for 3 times until no protein appears in the organic solvent in the lower layer;
and (3) dialysis: concentrating the protein-removed polysaccharide solution at low temperature under reduced pressure, placing into a pretreated dialysis bag (specification MD34, molecular weight cutoff is 3500 Da), placing into a large-capacity beaker, adding distilled water, standing for dialysis, changing water every 8h, and dialyzing for 24h. By 1% AgNO 3 Checking the residual conditions of NaCl, KCl and the like;
94.6g of red clover powder is extracted according to the embodiment 1, protein is removed by the method of the embodiment 4, and the purified polysaccharide is finally obtained by dialysis, wherein the extraction rate is 0.83g, and the extraction rate is 0.877 percent.
Example 4
DPPH belongs to a synthetic stable free radical, has characteristic absorption in a visible light region, and is simple, convenient, rapid and good in reproducibility.
The flavonoid and flavone and polysaccharide mixed samples prepared in examples 1-3 were all prepared with 70% ethanol, the polysaccharide sample, the positive control vitamin C (Vc) and vitamin E (Ve) were prepared with distilled water, and each test substance was prepared into 1.280mg/mL stock solution, which was then diluted to 7 concentration gradients for use.
Accurately adding 75 mu L of solution to be detected with each concentration into a 96-well enzyme label plate, adding 150 mu L of 0.1mM DPPH 70% ethanol solution, setting blank controls in reaction wells with each concentration, wherein the blank controls comprise 75 mu L of solution to be detected and 150 mu L of 70% ethanol, the total reaction volume is 225 mu L, fully shaking up, standing in the dark at room temperature for 30min, and measuring the light absorption value at the position of 519 nm. A decrease in absorbance indicates an increase in free radical scavenging activity. Each sample was replicated 3 times per concentration.
Clearance = [ a = 0 -(A 1 -A 2 )]/A 0
A0 is the light absorption value of DPPH solution with zero concentration of the solution to be detected, A1 is the light absorption value of the solution to be detected after the reaction with DPPH, and A2 is a blank control, namely the light absorption value of each solution to be detected and a blank solvent.
TABLE 1 DPPH antioxidant Activity
Note: p represents refined red clover polysaccharide, IF represents refined red clover isoflavone, and the two are mixed according to the mass ratio. Lower case letters indicate significant differences (P < 0.05).
As can be seen from Table 1, the activity of the purified polysaccharide from Trifolium pratense is comparable to that of the purified isoflavone. The mixing mass ratio of the two is 1: the activity is highest when 2, and is obviously better than that of each single product. DPPH activity decreased significantly with increasing polysaccharide content.
Example 5
Determination of superoxide anion antioxidant Activity
O 2- A type of radical having a long life in the organism. In a free radical chain reaction, O 2- It is used as initiator to promote the generation of other free radicals, further harms the organism and is closely related to various diseases. Therefore, the sample is typically purged of O 2- The ability of (A) to act as an important indicator of the activity of the antioxidant substance.
The flavonoid and flavone and polysaccharide mixed samples prepared in examples 1-3 are all prepared by 70% ethanol, the polysaccharide sample, the positive control vitamin C (Vc) and the vitamin E (Ve) are prepared by distilled water, each substance to be tested is prepared into 1.280mg/mL stock solution, and then diluted to 7 concentration gradients for standby.
Superoxide anion (O) scavenging determination by PMS-NADH-NBT system 2- Active by reducing Phenazine Methosulfate (PMS) with reduced coenzyme I (NADH) to produce superoxide anion, O 2- Reacting with nitrotetrazolium chloride (NBT) to generate a blue product with a strong absorption peak at 560nm, and inhibiting O after an antioxidant substance is added into a PMS-NADH-NBT system 2- The ability of the antioxidant substance to scavenge free radicals can be calculated by measuring the absorbance in the reaction with NBT. To a 96-well plate, 30. Mu.L of NBT (144. Mu. Mol/L), 100. Mu.L of a solution to be tested (2.50-125.0. Mu.g/mL, blank with a solvent instead of the sample solution), 30. Mu.L of NADH (0.68 mmol/L, prepared with phosphate buffer), 30. Mu.L of freshly prepared PMS (60. Mu. Mol/L), 30. Mu.L of pH 7.4 phosphate buffer were sequentially added. The blank tube is 120 mu L of buffer solution and 100 mu L of solution to be detected, and the light absorption value of the solution at 560nm is determined after oscillation for 30s and 5 min.
TABLE 2 superoxide anion antioxidant Activity
Note: p represents refined red clover polysaccharide, IF represents refined red clover isoflavone, and the two are mixed according to the mass ratio. Lower case letters indicate significant differences (P < 0.05).
As can be seen from Table 2, the activity of isoflavone of red clover is equivalent to Vc and Ve, while the activity of polysaccharide of red clover is obviously higher than Vc and Ve, especially the activity of refined polysaccharide is 100 times higher than that of positive control. The activity of the 3 compounds is equivalent to that of refined polysaccharide, and the activity is slightly enhanced along with the increase of the content of the polysaccharide.
Example 6
The effect of the combined antioxidation of the red clover isoflavone and the polysaccharide is evaluated by utilizing a joint index (CI) which is a method for judging the relationship between the antioxidant activities of the two components. We here used the median effect principle of Talady and Chou in combination with the index of action CI.
CI＝D 1 /D X1 +D 2 /D X2
D 1 And D 2 Is the action concentration of the flavone and the polysaccharide when the combined action inhibition rate is 50 percent. D x1 And D x2 Is the action concentration of flavone and polysaccharide with the action inhibition rate of 50% when acting alone.
According to the theory of Talady and Chou, if CI =1, it indicates that the combined effect of drug 1 and drug 2 is additive; when CI is greater than 1, the combined effect of the drug 1 and the drug 2 is shown to be antagonistic; if CI <1, it indicates that drug 1 and drug 2 act in combination synergistically.
TABLE 3 Combined antioxidation index CI
Note: lower case letters indicate significant differences (P < 0.05).
As can be seen from table 3, the mixing mass ratio of polysaccharide to isoflavone was 1:2, the two components are synergistic, wherein the synergistic effect on the superoxide anion is stronger. However, as the polysaccharide content increases, a degree of antagonism of the two components occurs with respect to DPPH activity.
Example 7
The water solubility determination method comprises the following steps: each measurement sample was accurately weighed at 5.0mg each, and 5.0mg was sequentially added to a transparent centrifuge tube containing 1mL of distilled water at 25 ℃ at room temperature until precipitation occurred. If the solution is not completely dissolved after the addition, the solution can be accelerated by a vortex oscillator, and then the amount of each sample added in 1mL of distilled water is recorded, and the mass of the undissolved part is observed and estimated to obtain the actual dissolved amount in 1mL of distilled water.
Table 4 dissolution ranges of different preparation ingredients in water
As is clear from Table 4, the solubility of the polysaccharide in water was higher than that of the isoflavone, and the solubility of the purified sample was improved. In particular, the solubility of the refined red clover polysaccharide (more than 25 g/L) is 5 times of that of the refined isoflavone and the crude polysaccharide.
The invention takes red clover as raw material, the functions and the application range of the prepared different types of natural antioxidant ingredients are different, which provides conditions for the application of multiple aspects and multiple purposes in the research and development and the production of cosmetics, and the market prospect is wide. Among them, the refined red clover polysaccharide has the advantages of high activity and good water solubility, and is considered to be adopted firstly. Secondly, crude red clover polysaccharide or red clover isoflavone may be considered if it is desired to prepare fat-soluble components. The two components have different activity, and crude red clover polysaccharide has high antioxidant activity to superoxide anion, while isoflavone has better DPPH activity. Thirdly, if the factors such as cost, comprehensive activity and solubility are considered, the two components can be adopted to carry out a compounding scheme, and particularly, the mass ratio of the polysaccharide to the isoflavone is 1:2, the synergistic effect between the two components is the best, and the activity is the highest.
In addition, the red clover is excellent pasture, and the isoflavone is also used as a health care product for marketing at home and abroad, so the cosmetics prepared by the plant are green and environment-friendly, are very safe to human bodies, and have rich resources.
In summary, the following steps: the red clover disclosed by the invention is leguminous perennial pasture, is widely planted in the global range and is rich in resources. The plant is reported to be rich in isoflavone compounds and has better antioxidant capacity. However, the antioxidant component extracted and prepared from the plant has better activity than the isoflavone component, better water solubility, wider application range and very good development and application prospect.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (3)
1. A natural antioxidant for cosmetics is characterized in that the main component is an extract of red clover extract, the component is a compound of refined polysaccharide and refined isoflavone, and the mass ratio of the compound of the refined polysaccharide to the refined isoflavone is 1:10-15:1, preparation.
2. A method for preparing a natural antioxidant for cosmetics according to claim 1, comprising the steps of:
s1: preparation of red clover extract
Deactivating enzyme of collected red clover leaves at 105 deg.C for 2-60min, drying at 65 deg.C for 1-160h, pulverizing, defatting with small amount of petroleum ether for 1-10 times, volatilizing solvent, air drying, weighing certain mass of defatted red clover powder, ultrasonic extracting with 10-50 times of 10-100% ethanol at 30-70 deg.C for 10-90min, filtering, collecting supernatant, extracting sample residue with 5-120 times of solvent for 10-90min, and mixing the two extractive solutions;
s2: preparation of crude polysaccharide and crude isoflavone
Crude polysaccharide: concentrating the extractive solution at 15-90 deg.C with vacuum rotary instrument until the volume is one eighth, adding 2-20 times volume of anhydrous ethanol into the concentrated solution under stirring, standing in 4 deg.C refrigerator overnight, centrifuging at 4800rpm for 2-20min, centrifuging to obtain brown precipitate as crude polysaccharide part, repeatedly washing the precipitate with anhydrous ethanol for 1-10 times, washing to remove alcohol soluble impurities, washing the precipitate with ethanol, pouring into culture dish, and volatilizing solvent to obtain brown red clover crude polysaccharide;
the supernatant is the isoflavone part, and the supernatant is decompressed and concentrated to obtain crude isoflavone of the red clover;
s3: refining of red clover isoflavone
Soaking the pretreated resin in distilled water for more than 10-90h, loading the column resin in wet method at a height of 3-50cm, dissolving 1-30g of crude isoflavone in water, loading the sample, waiting for adsorption for 0.5-24h, eluting with distilled water, 30% and 50% ethanol for 1-5BV respectively, eluting with 70% ethanol for 1-5BV, finally eluting with 100% ethanol for 1-10BV, collecting 70% ethanol eluate, and concentrating under reduced pressure to obtain refined isoflavone;
s4: refining of red clover polysaccharide
Protein removal: preparing 1-10mg/mL solution of red clover crude polysaccharide with distilled water, adding 1/4 volume of sevage solution, oscillating for 1-30min, transferring to a separating funnel, standing for 1-120min, discharging the lower layer solution and the middle layer protein, and repeating the operation for 1-6 times until the protein does not appear in the lower layer organic solvent;
and (3) dialysis: concentrating the protein-removed polysaccharide solution at low temperature under reduced pressure, filling into a pretreated dialysis bag, placing into a large-capacity beaker, filling distilled water, standing for dialysis, changing water every 1-16h, dialyzing for 1-72h, and checking residual conditions of NaCl, KCl and the like by using 1% AgNO3 to finally obtain the purified polysaccharide.
3. The method for preparing a natural antioxidant for cosmetics according to claim 2, wherein, for the step S3, the resin pretreatment: soaking the resin in 20-100% ethanol for 1-72h, filtering, repeatedly washing with deionized water until there is no alcohol smell and the washing solution has no white turbidity, soaking in 1-30% NaOH for 1-72h, filtering, repeatedly washing the resin with deionized water until pH is neutral, soaking in 1-30% HCl solution for 1-72h, filtering and washing the resin to neutral, and drying at 20-90 deg.C for use.
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