CN111978420A - Preparation method of spirulina selenium polysaccharide - Google Patents
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Abstract
The invention discloses a preparation method of spirulina selenium polysaccharide, which comprises the following steps: s1, pretreatment of spirulina powder: s2, protein separation: s3: purifying the crude polysaccharide: s4: and (3) carrying out sulfation reaction: s5: and (4) carrying out selenic acid esterification reaction to obtain the product. According to the preparation method, the spirulina powder is subjected to ball-milling pretreatment, impurities in the spirulina powder can be removed to the maximum extent, the yield of the polysaccharide is improved, and the yield of the spirulina selenium polysaccharide is improved by performing catalytic sulfonation reaction to reduce substitution of hydroxyl groups and generation of impurities.
Description
Technical Field
The invention relates to the field of selenium polysaccharide preparation, in particular to a preparation method of spirulina selenium polysaccharide.
Background
Selenium element is a trace element necessary for human body, while inorganic selenium has toxicity and small safe dose, organic selenide is a novel compound, and has pharmacological activities of resisting virus, resisting tumor, resisting inflammation, resisting aging, promoting growth, protecting visual organs, detoxifying, preventing and treating cardiovascular diseases and preventing liver diseases, etc., selenium has unique biochemical property and pharmacological action, and the development of selenium-containing compounds is very significant. Selenium polysaccharide is an organic selenium compound that has both selenium and polysaccharide activities through the combination of polysaccharide and selenium. The biological activity of the selenium polysaccharide is generally higher than that of selenium and polysaccharide, and the selenium polysaccharide is easier to be absorbed and utilized by organisms, so the selenium polysaccharide has wide application in the aspects of immunoregulation, tumor resistance, oxidation resistance, aging resistance and the like.
The content of selenium polysaccharide in plants or microorganisms is low, and at present, people mainly obtain the high content of selenium polysaccharide by the following two methods: artificial selenium treatment is carried out to obtain selenium-rich plants so as to form natural selenium polysaccharide and obtain artificial selenium polysaccharide by utilizing polysaccharide selenylation. The artificial selenium polysaccharide is prepared by introducing sulfate groups into polysaccharide molecules through sulfonation reaction and the like of partial plant polysaccharides without sulfate radicals in nature, and then substituting partial sulfate radicals in the polysaccharide molecules with selenite radicals in sodium selenite to form selenate lipopolysaccharide. The natural selenium polysaccharide has less amount and high price, which is not beneficial to industrial production, and the technology of artificially synthesizing the selenium polysaccharide is complex and has low selenium content.
The above background disclosure is only for the purpose of assisting understanding of the inventive concept and technical solutions of the present invention, and does not necessarily belong to the prior art of the present patent application, and should not be used for evaluating the novelty and inventive step of the present application in the case that there is no clear evidence that the above content is disclosed at the filing date of the present patent application.
Disclosure of Invention
Technical problem to be solved
The invention aims to provide a preparation process of selenium-rich spirulina selenium polysaccharide, which aims to solve the technical problems of low yield, high impurity content and multiple purification of the spirulina selenium polysaccharide in the prior art.
(II) technical scheme
In order to achieve the purpose, the invention provides the following technical scheme:
a preparation method of spirulina selenium polysaccharide comprises the following steps:
s1, pretreatment: drying spirulina powder at 50 deg.C for 8-10h, cooling to room temperature, and ball milling at 30-35 deg.C for 5-7 days;
s2, protein separation: dissolving the spirulina powder treated in the step S1 and water according to the weight ratio of 1:1-2, adding the mixed solution, fully stirring, centrifuging for 20-30min at 3000-4000r/min, removing protein on a two-phase interface, collecting upper-layer filtrate, repeatedly adding the mixed solution for 5-6 times of centrifugal washing operation, finally precipitating the obtained upper-layer filtrate with absolute ethyl alcohol until the alcohol content is 80%, standing for 24h at 4 ℃, centrifuging, collecting precipitate, and vacuum drying to obtain crude spirulina polysaccharide;
s3, purifying polysaccharide: washing the crude spirulina polysaccharide obtained in the step S2 with purified water for 3-4 times, centrifuging for 10-15min at 10000-12000r/min, collecting precipitate, and vacuum drying to obtain purified polysaccharide;
s4, sulfation reaction: cooling pyridine to-10 to-5 ℃, slowly adding chlorosulfonic acid, then adding the purified polysaccharide obtained in the step S3, preserving heat for 1-1.5 hours at the temperature of 80-90 ℃, cooling to room temperature, adding ice water, then adding ethanol, precipitating out, filtering to obtain a first filter cake, pulping the filter cake in 50% alcohol solution, heating, cooling to-5 to 0 ℃, filtering to obtain a second filter cake, and drying at the temperature of 45-50 ℃ to obtain spirulina polysaccharide sulfate;
s5, esterification of selenic acid: adding the spirulina polysaccharide sulfate obtained in the step S4 into mixed acid, adding a selenizing reagent and an auxiliary agent, and reacting for 6-7h at 60-70 ℃ under the protection of N2; and carrying out post-treatment to obtain the spirulina selenium polysaccharide.
Preferably, in step S5, the mixed acid includes nitric acid with a concentration of 0.6% and perchloric acid with a concentration of 0.2%.
Preferably, in step S5, the selenizing reagent is one of sodium selenite and potassium selenite.
Preferably, in step S5, the auxiliary is barium chloride.
Preferably, in step S5, the post-treatment process is to cool the material to 20-25 ℃ under the protection of N2 after the reaction is finished, add sodium carbonate, adjust the pH to 6-7, filter to obtain a primary filtrate, add an appropriate amount of anhydrous sodium sulfate into the primary filtrate, stir, and then stand for 1-2h to filter to obtain a secondary filtrate.
Preferably, in step S5, the secondary filtrate is filled into a dialysis bag and dialyzed with running water overnight, then a small amount of dialysate is added with ascorbic acid for detection, dialysis is stopped until no red color is formed, then the dialysate is distilled under reduced pressure to be thick paste, precipitated with absolute ethyl alcohol, placed in a refrigerator overnight, filtered, and the precipitate is washed with absolute ethyl alcohol and dried in vacuum.
(III) advantageous effects
Compared with the prior art, the spirulina selenium polysaccharide has the following advantages:
according to the method, the spirulina powder is subjected to ball-milling pretreatment, impurities in the spirulina powder can be removed to the maximum extent, the yield of polysaccharide is improved, the catalytic sulfonation reaction is carried out, the substitution of hydroxyl groups is reduced, the generation of impurities is reduced, the yield of spirulina selenium polysaccharide is improved, the selenium content can reach more than 52g/Kg, and no potassium ions, sodium ions, barium ions and the like are left.
Drawings
FIG. 1 is the chemical structural formula of the spirulina selenium polysaccharide.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. 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.
FIG. 1 shows the spirulina selenium polysaccharide prepared by the present invention.
Example 1
A preparation method of spirulina selenium polysaccharide comprises the following steps:
s1, pretreatment: drying 1kg of spirulina powder at 50 ℃ for 8h, cooling to room temperature, and ball-milling at 30 ℃ for 5 days;
s2, protein separation: mixing and dissolving the spirulina powder treated in the step S1 with 1kg of water, adding 5L of mixed solution (the mixed solution is obtained by uniformly mixing chloroform: n-butanol according to a volume ratio of 4: 1), fully stirring for 30min, centrifuging at 3000r/min for 20min, removing protein on a two-phase interface, and collecting upper-layer filtrate; adding 5L of mixed solution again for centrifugal washing, namely adding the mixed solution, fully stirring for 30min, centrifuging for 30min at 3000r/min, removing protein on a two-phase interface, and collecting upper filtrate; repeatedly adding 5L of mixed solution into the upper layer filtrate, centrifuging and washing for 4 times, precipitating the finally obtained upper layer filtrate with absolute ethanol until the alcohol content is 80%, standing for 24h at 4 ℃, centrifuging and collecting precipitate, and drying in vacuum to obtain 100-fold 105g of crude spirulina polysaccharide;
s3: purifying polysaccharide: washing the crude Spirulina polysaccharide obtained in step S2 with 300ml of purified water at 2 deg.C, repeating the above purified water washing for 3 times, centrifuging at 10000r/min for 10min, collecting precipitate, and vacuum drying to obtain 50g of purified polysaccharide;
s4: and (3) carrying out sulfation reaction: cooling 100ml of pyridine to-10 ℃, slowly adding 4ml of chlorosulfonic acid (the content is more than or equal to 85%), then adding the purified polysaccharide obtained in the step S3, preserving heat for 1h at 80 ℃, cooling to room temperature, adding 15g of ice water, then adding 30ml of ethanol, precipitating, filtering to obtain a first filter cake, pulping the filter cake in 50ml of 50% alcohol solution, heating, cooling to-5 ℃, filtering to obtain a second filter cake, and drying at 45 ℃ to obtain 36g of spirulina polysaccharide sulfate;
s5: and (3) selenic acid esterification reaction: adding the spirulina polysaccharide sulfate obtained in the step S4 into 100g of mixed acid, wherein the mixed acid comprises 0.6% of nitric acid and 0.2% of perchloric acid, adding 1g of sodium selenite and 0.2g of barium chloride, stirring uniformly, and adding into N2Reacting for 6 hours at 60 ℃ under the protection condition; then carrying out post-treatment, wherein the post-treatment process is that after the reaction is finished, under the protection of N2, the material is cooled to 20 ℃, sodium carbonate is added, the pH value is adjusted to 6, the filtration is carried out to obtain a primary filtrate, a proper amount of anhydrous sodium sulfate is added into the primary filtrate, the primary filtrate is stirred and then is kept stand for 1h, and the secondary filtrate is obtained; putting the secondary filtrate into dialysis bag, dialyzing with flowing water overnight, collecting a small amount of dialysate, adding ascorbic acid, detecting, stopping dialysis until no red color appears, distilling under reduced pressure to obtain soft extract,precipitating with anhydrous ethanol, standing in a refrigerator overnight, filtering, washing the precipitate with anhydrous ethanol, and vacuum drying to obtain the spirulina selenium polysaccharide 20 g.
Example 2
A preparation method of spirulina selenium polysaccharide comprises the following steps:
s1, pretreatment: drying 1kg of spirulina powder at 50 ℃ for 10h, cooling to room temperature, and ball-milling at 35 ℃ for 7 days;
s2, protein separation: mixing and dissolving the spirulina powder treated in the step S1 with 2kg of water, adding 5L of mixed solution (the mixed solution is obtained by uniformly mixing chloroform: n-butanol according to a volume ratio of 4: 1), fully stirring for 40min, centrifuging for 30min at 4000r/min, removing protein on a two-phase interface, and collecting upper-layer filtrate; adding 5L of mixed solution again for centrifugal washing, namely adding the mixed solution, fully stirring for 40min, centrifuging for 30min at 4000r/min, removing protein on a two-phase interface, and collecting upper filtrate; repeatedly adding 5L of mixed solution into the upper layer filtrate, centrifuging and washing for 3 times, precipitating the final upper layer filtrate with anhydrous ethanol until the ethanol content is 80%, standing at 4 deg.C for 24 hr, centrifuging, collecting precipitate, and vacuum drying to obtain 105g crude Spirulina polysaccharide;
s3: purifying polysaccharide: washing the crude Spirulina polysaccharide obtained in step S2 with 300ml of purified water at 4 deg.C, repeating the above washing with purified water for 3 times, centrifuging at 12000r/min for 15min, collecting precipitate, and vacuum drying to obtain 54g of purified polysaccharide;
s4: and (3) carrying out sulfation reaction: cooling 100ml of pyridine to-5 ℃, slowly adding 4ml of chlorosulfonic acid (the content is more than or equal to 85%), then adding the purified polysaccharide obtained in the step S3, preserving heat for 1.5h at 90 ℃, cooling to room temperature, adding 15g of ice water, then adding 30ml of ethanol, precipitating and filtering to obtain a first filter cake, pulping the filter cake in 50ml of 50% alcohol solution, heating, cooling to-5-0 ℃, filtering to obtain a second filter cake, and drying at 50 ℃ to obtain 39g of spirulina polysaccharide sulfate;
s5: and (3) selenic acid esterification reaction: adding the spirulina polysaccharide sulfate obtained in step S4 into 100g of mixed acid containing nitric acid with concentration of 0.6% and perchloric acid concentration of 0.2%, adding 1g sodium selenite or potassium selenite and 0.2g barium chloride, stirring, and adding N2Reacting for 7 hours at 70 ℃ under the protection condition; then carrying out post-treatment, wherein the post-treatment process is that after the reaction is finished, under the protection of N2, the material is cooled to 25 ℃, sodium carbonate is added, the pH value is adjusted to 7, the filtration is carried out to obtain a primary filtrate, a proper amount of anhydrous sodium sulfate is added into the primary filtrate, the primary filtrate is stirred and then is kept stand for 2 hours, and the secondary filtrate is obtained; and (3) putting the secondary filtrate into a dialysis bag, dialyzing with running water overnight, taking a small amount of dialysate, adding ascorbic acid for detection, stopping dialysis until no red color exists, distilling under reduced pressure to obtain thick paste, precipitating with absolute ethyl alcohol, standing in a refrigerator overnight, washing the precipitate with absolute ethyl alcohol after suction filtration, and drying in vacuum to obtain 22g of spirulina selenium polysaccharide.
Example 3
A preparation method of spirulina selenium polysaccharide comprises the following steps:
s1, pretreatment: drying 1kg of spirulina powder at 50 ℃ for 9h, cooling to room temperature, and ball-milling at 30-35 ℃ for 6 days;
s2, protein separation: mixing and dissolving the spirulina powder treated in the step S1 with 1.5kg of water, adding 5L of mixed solution (the mixed solution is obtained by uniformly mixing chloroform: n-butanol according to a volume ratio of 4: 1), fully stirring for 40min, centrifuging at 3500r/min for 25min, removing protein on a two-phase interface, and collecting upper-layer filtrate; adding 5L of mixed solution again, centrifuging, i.e. adding mixed solution, stirring thoroughly for 35min, centrifuging at 3500r/min for 25min, removing protein at two-phase interface, and collecting upper filtrate; repeatedly adding 5L of mixed solution into the upper layer filtrate, centrifuging and washing for 4 times, precipitating the final upper layer filtrate with anhydrous ethanol until the ethanol content is 80%, standing at 4 deg.C for 24 hr, centrifuging, collecting precipitate, and vacuum drying to obtain crude Spirulina polysaccharide 102 g;
s3: purifying polysaccharide: washing the crude Spirulina polysaccharide obtained in step S2 with 300ml of purified water at 3 deg.C, repeating the above purified water washing for 3 times, centrifuging at 11000r/min for 12min, collecting precipitate, and vacuum drying to obtain 52g of purified polysaccharide;
s4: and (3) carrying out sulfation reaction: cooling 100ml of pyridine to-8 ℃, slowly adding 4ml of chlorosulfonic acid (the content is more than or equal to 85%), then adding the purified polysaccharide obtained in the step S3, preserving heat for 1.2h at 85 ℃, cooling to room temperature, adding 15g of ice water, then adding 30ml of ethanol, precipitating and filtering to obtain a first filter cake, pulping the filter cake in 50ml of 50% alcohol solution, heating, cooling to-2 ℃, filtering to obtain a second filter cake, and drying at 47 ℃ to obtain 37g of spirulina polysaccharide sulfate;
s5: and (3) selenic acid esterification reaction: adding the spirulina polysaccharide sulfate obtained in step S4 into 100g of mixed acid containing 0.6% nitric acid and 0.2% perchloric acid, adding 1g of sodium selenite or potassium selenite and 0.2g of barium chloride, stirring, and adding into N2Reacting for 6.5 hours at 65 ℃ under the protection condition; then carrying out post-treatment, wherein the post-treatment process is that after the reaction is finished, under the protection of N2, the material is cooled to 23 ℃, sodium carbonate is added, the pH value is adjusted to 6.5, the mixture is filtered to obtain a primary filtrate, a proper amount of anhydrous sodium sulfate is added into the primary filtrate, and the primary filtrate is stirred and then stands for 1.5h for filtering to obtain a secondary filtrate; and (3) putting the secondary filtrate into a dialysis bag, dialyzing with running water overnight, taking a small amount of dialysate, adding ascorbic acid for detection, stopping dialysis until no red color exists, distilling under reduced pressure to obtain thick paste, precipitating with absolute ethyl alcohol, standing in a refrigerator overnight, washing the precipitate with absolute ethyl alcohol after suction filtration, and drying in vacuum to obtain 20.5g of the spirulina selenium polysaccharide.
Comparative example 1
Step S4 is not present, and the preparation of spirulina selenium polysaccharide is carried out under the same conditions as in example 2;
comparative example 2
In step S5, the spirulina selenium polysaccharide is prepared without adding perchloric acid and under the same conditions as those in example 2;
comparative example 3
In step S5, the spirulina selenium polysaccharide is prepared without adding perchloric acid, the concentration of the dilute nitric acid is 3 percent, and the other conditions are the same as those in the example 2;
comparative example 4
In step S5, the secondary filtrate is dialyzed overnight without flowing water, directly distilled under reduced pressure to obtain thick paste, precipitated with anhydrous ethanol, placed in a refrigerator overnight, filtered, washed with anhydrous ethanol, and vacuum dried.
The related tests of the spirulina selenium polysaccharide prepared in the examples 1 to 3 and the comparative examples 1 to 4 are as follows:
1. detection of selenium content using spectrophotometry
Taking 0.00, 1.25, 2.50, 5.00, 10.00, 15.00, 20.00 and 25.00mL of 2 mu g/mL selenium standard solution, respectively placing the solution in a 125mL separating funnel, adding 30mL of water, adjusting the pH value to 2 by using a dilute hydrochloric acid solution, adding 3.0mL of 1% o-phenylenediamine hydrochloride test solution, shaking the solution uniformly, placing the solution for 2 hours, then adding 10.00mL of toluene, extracting, standing the solution, taking a toluene layer, taking 0mL of selenium-free solution as a blank control solution, performing spectral scanning in a 190-800 nm range of an ultraviolet spectrophotometer, and determining the maximum absorption wavelength to be 338 nm. And (3) measuring the absorbance value A at the maximum absorption wavelength, drawing a standard curve by taking the selenium content (g) as a horizontal coordinate and the absorbance A as a vertical coordinate, and solving a regression equation. 0.01g of the finished product of the examples and comparative examples was ground and put into a 100mL conical flask, and 4mL of mixed acid (V (HClO) was added4):V(H2SO4):V(HNO3) 1:1:4) and left overnight. The flask was then placed in a thermostatted water bath and allowed to stand at 95 ℃ for about 1.5 h. And then dividing the solution into two parts, transferring one part into a separating funnel, processing according to the method for drawing the standard curve, measuring absorbance by taking 338nm as a detection wavelength, and calculating the selenium content in the sample according to a regression equation.
The results of the selenium contents according to the above method for examples and comparative examples are shown in the following table 1:
TABLE 1
| Group of | Example 1 | Example 2 | Example 3 | Comparative example 1 | Comparative example 2 | Comparative example 3 | Comparative example 4 |
| Selenium content (g/kg) | 52 | 63 | 57 | 0.5 | 30 | 15 | 70 |
| Final product yield (%) | 2.0 | 2.2 | 2.1 | 3.0 | 2.1 | 1.0 | 2.5 |
From the above table 1, in terms of selenium content and yield, it can be seen that in examples 1-3, the selenium content is greater than or equal to 52g/kg, and the final product yield is 2.0% -2.2%; in contrast, in comparative example 1, the product was prepared with a selenium content of only 0.5g/kg, which is greatly reduced, indicating that purification was performed in the absence of step S4Adding polysaccharide into mixed acid (0.6% nitric acid and 0.2% perchloric acid), adding potassium selenite and barium chloride, and adding N2Under the protection condition, reacting for 6.5h at 66 ℃ to perform direct esterification reaction, wherein the selectivity of the reaction between potassium selenite and purified polysaccharide is poor, so that the selenium content in the spirulina selenium polysaccharide is low; in contrast, in comparative example 2, the selenium content in the spirulina selenium polysaccharide prepared in the absence of perchloric acid is reduced, which is supposed to be that step S5 of example 2 is more acidic in a mixed solution of perchloric acid and nitric acid, and the spirulina polysaccharide sulfate is promoted by barium chloride to more easily generate spirulina selenium polysaccharide (esterified substance); in the comparative example 3, the concentration of the nitric acid is 3%, and the concentration of the nitric acid is relatively higher than that of the nitric acid in the example 2, so that the oxidability is relatively higher than that of the example 2, the spirulina polysaccharide can be oxidized, the product is oxidized, the selenization reaction is influenced, and the product yield and the selenium content are obviously reduced; in comparative example 4, in which the post-treatment without dialysis was not performed, the selenium content and the yield were increased to some extent from example 2, which is likely to be the end product containing selenized residual selenate, barium salt, potassium salt, sodium salt, etc. in step S5.
2. Flame test
Platinum wire, alcohol burner, dilute hydrochloric acid, blue cobalt glass.
Procedure of operation
Firstly, burning a platinum wire dipped with diluted hydrochloric acid on a colorless flame until the platinum wire is colorless; ② dipping samples to burn on colorless flame to observe flame color (observing through blue cobalt glass)
| Group of | Example 1 | Example 2 | Example 3 | Comparative example 4 |
| Direct flame color | Colorless and colorless | Colorless and colorless | Colorless and colorless | Yellow green |
| Through blue cobalt glass | Colorless and colorless | Colorless and colorless | Colorless and colorless | Yellow green |
Remarking: the color of Ba flame is yellow-green; the Na flame color is yellow, and the Ka flame color is yellow-green;
as can be seen from examples 1-3, the direct flame color and the flame color observed through blue cobalt glass are colorless, which indicates that the prepared spirulina selenium polysaccharide has no sodium, potassium and barium residues and does not bring the risk of barium ion poisoning; in contrast, the direct flame color of the comparative example 1 and the yellow-green flame color observed through blue cobalt glass indicate that barium remains and the risk of barium ion poisoning exists in the product; therefore, it can be said that in the post-treatment of step S5, excessive selenite is effectively removed by dialysis and inorganic compounds (sodium, potassium, barium salts, etc.) are dialyzed away.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that 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 (6)
1. A preparation method of spirulina selenium polysaccharide is characterized by comprising the following steps: the method comprises the following steps:
s1, pretreatment: drying spirulina powder at 50 deg.C for 8-10h, cooling to room temperature, and ball milling at 30-35 deg.C for 5-7 days;
s2, protein separation: dissolving the spirulina powder treated in the step S1 and water according to the weight ratio of 1:1-2, adding the mixed solution, fully stirring, centrifuging for 20-30min at 3000-4000r/min, removing protein on a two-phase interface, collecting upper-layer filtrate, repeatedly adding the mixed solution for 5-6 times of centrifugal washing operation, finally precipitating the obtained upper-layer filtrate with absolute ethyl alcohol until the alcohol content is 80%, standing for 24h at 4 ℃, centrifuging, collecting precipitate, and vacuum drying to obtain crude spirulina polysaccharide;
s3, purifying polysaccharide: washing the crude spirulina polysaccharide obtained in the step S2 with purified water for 3-4 times, centrifuging for 10-15min at 10000-;
s4, sulfation reaction: cooling pyridine to-10 to-5 ℃, slowly adding chlorosulfonic acid, then adding the purified polysaccharide obtained in the step S3, preserving heat for 1-1.5 hours at the temperature of 80-90 ℃, cooling to room temperature, adding ice water, then adding ethanol, precipitating out, filtering to obtain a first filter cake, pulping the filter cake in 50% alcohol solution, heating, cooling to-5 to 0 ℃, filtering to obtain a second filter cake, and drying at the temperature of 45-50 ℃ to obtain spirulina polysaccharide sulfate;
s5, esterification of selenic acid: adding the spirulina polysaccharide sulfate obtained in the step S4 into mixed acid, adding a selenizing reagent and an auxiliary agent into the mixed acid, and adding N2Reacting for 6-7h at 60-70 ℃ under the protection condition; and carrying out post-treatment to obtain the spirulina selenium polysaccharide.
2. The method for preparing spirulina selenium polysaccharide of claim 1, which is characterized in that: in the step S5, the mixed acid includes nitric acid with a concentration of 0.6% and perchloric acid with a concentration of 0.2%.
3. The method for preparing spirulina selenium polysaccharide of claim 1, which is characterized in that: in step S5, the selenizing reagent is one of sodium selenite and potassium selenite.
4. The method for preparing spirulina selenium polysaccharide of claim 1, which is characterized in that: in the step S5, the auxiliary agent is barium chloride.
5. The method for preparing spirulina selenium polysaccharide of claim 1, which is characterized in that: in the step S5, the post-treatment process is performed after the reaction is finished and N is2Under the protection condition, cooling the materials to 20-25 ℃, adding sodium carbonate, adjusting the pH value to 6-7, filtering to obtain a primary filtrate, adding a proper amount of anhydrous sodium sulfate into the primary filtrate, stirring, standing for 1-2h, and filtering to obtain a secondary filtrate.
6. The method of claim 5, wherein the method comprises the steps of: and in the step S5, putting the secondary filtrate into a dialysis bag, dialyzing with running water overnight, taking a small amount of dialysate, adding ascorbic acid for detection, stopping dialysis until no red color exists, distilling under reduced pressure to obtain thick paste, precipitating with absolute ethyl alcohol, standing in a refrigerator overnight, filtering, washing the precipitate with absolute ethyl alcohol, and vacuum drying.
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| CN113951367A (en) * | 2021-11-16 | 2022-01-21 | 安徽中志科技有限公司 | Edible fungus protein powder with high organic selenium content, preparation method and application |
| CN115417935A (en) * | 2022-09-22 | 2022-12-02 | 潘惟谦 | Preparation method of seaweed selenium polysaccharide |
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| CN115417935A (en) * | 2022-09-22 | 2022-12-02 | 潘惟谦 | Preparation method of seaweed selenium polysaccharide |
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