CN106832030A - The method that active polysaccharide is extracted from marine alga - Google Patents
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- CN106832030A CN106832030A CN201710046367.0A CN201710046367A CN106832030A CN 106832030 A CN106832030 A CN 106832030A CN 201710046367 A CN201710046367 A CN 201710046367A CN 106832030 A CN106832030 A CN 106832030A
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
Abstract
The present invention provides a kind of method that active polysaccharide is extracted from marine alga, and operating procedure is:1)Degreasing, de- glycosides;2)Enzymolysis;3)Deproteinized;4)Ultrafiltration;5)Gel chromatography;6)Desalt, dry.Have the beneficial effect that:With acetone petroleum ether mixed-liquor return degreasing, glycoside and alkaloid are taken off with alcohol reflux, carry out removal of impurities;Complex enzyme formulation digests the efficiency high of cellulose and pectin, and digests fully, and the destructive rate to cell membrane is high;It is aided with ultrasonic wave extraction, active polysaccharide flows out from alginic cell and is dissolved in the water, and the recovery rate of active polysaccharide is high;Rough marine alga active polysaccharide powder is isolated and purified using ultrafiltration, gel chromatography and dialysis, the marine alga active polysaccharide purity for finally obtaining is very high, activity is strong, organic solvent-free residual, green health is adapted to be applied to cosmetics, medicine, field of food.
Description
Technical field
The present invention relates to technical field of polysaccharide extraction, specifically a kind of method that active polysaccharide is extracted from marine alga.
Background technology
In recent years, substantial amounts, miscellaneous marine organisms are in fields such as food, medicine, cosmetics and biomaterials
It is used widely, oneself turns into the focus of Natural products research to extract isolating biologically active material from marine organisms.Give birth to ocean
Thing most strikingly its abundant medicine resource, scientist has separated and has identified various marine naturals from marine organisms
Active material, many of which has the biological actions such as antibacterial, antiviral, antitumor and anticoagulation, to develop ocean
New drug lays the foundation.Compared with the other biological macromolecular such as protein, fat, nucleic acid, carbohydrate has stronger hydrophily.Ocean
, by synthesizing polysaccharose substance, this is special to adapt to ocean with the free moisture required for vital movement in keeping body for biology
Environment.Polysaccharide wide participation cell recognition, cell growth, differentiation, metabolism, embryonic development, cell carcinogenesis, virus infect and exempt from
The items vital movement such as epidemic disease response, with antiviral, anticoagulation, antitumor, anti-mutation, antiulcer, anti-oxidant, hypoglycemic and drop
The bioactivity such as blood fat, as modern medicine and the common focus of attention of functional food worker.Algal polysaccharides are contained by marine alga
Various polymeric carbohydrates, account for the 20 ~ 70% of dry weight, be the main component of marine alga.Since the sixties in 20th century, people
Gradually find that algal polysaccharides have many bioactivity, such as antiviral, antitumor, immunological regulation, promoting blood circulationization addiction, anti-ageing
Old and protection body cell etc., and it is most nontoxic, there are larger potentiality in terms of marine drug is developed into.
Now existing many reports on Polyose extraction, prior art such as Authorization Notice No. are in CN103497258B
State's patent of invention, discloses a kind of method for extraction and purification of active polysaccharide of wheat bran, and its step includes:Pretreatment, degreasing,
Polyose extraction and Thick many candies are refined, finally to the crude product for extracting and the yield of component, chemical composition and immunological regulation after purification
Activity is analyzed.The method for extraction and purification of the active polysaccharide of wheat bran, is that wheat bran is obtained using water extraction and alcohol precipitation method
Polysaccharide crude, recycles agarose Gel column purifying to obtain the polysaccharide with immunoregulatory activity, and its extracting method is simple, operation
Low cost, its yield of the wheat bran Thick many candies of gained is 5.0%, and by purifying active polysaccharide of wheat bran, its yield is
5.6%, and with stronger immunoregulatory activity.
Prior art such as Authorization Notice No. is the Chinese invention patent of CN103265644B, discloses a kind of larva of a silkworm with batrytis anticancer
The preparation method of active polysaccharide BBPW-2.Larva of a silkworm with batrytis powder removes glycosides through acetone-petroleum ether backflow degreasing and 80% alcohol reflux
After class and alkaloid, with distilled water in extracting Thick many candies at 80 ~ 100 DEG C.After Thick many candies remove isolating protein through Sevag methods,
Neutral polysaccharide component is isolated with DEAE Sepharose ion displacement chromatographies, then through Sephacryl S-300 layers
Analysis, takes the 2nd eluting peak, is further purified through Sephacryl S-200 chromatographies after being concentrated under reduced pressure, and freeze-drying is obtained
To larva of a silkworm with batrytis active anticancer Polysaccharide B BPW-2.Product prepared by the method, purity is homogeneous, and molecular weight is 2.0 × 10 3Da is right
The growth of human cervical carcinoma cell Hela and human liver cancer cell HepG2 has inhibitory action, to normal cell embryo nephrocyte
HEK293 and mouse macrophage RAW264.7 growths have no adverse effects, and can be used for antineoplastic product exploitation.Both approaches
The recovery rate of polysaccharide is low, and consumption of organic solvent is big, and the pollution to environment is big.
The content of the invention
High it is an object of the invention to provide a kind of polysaccharide extract rate, purity is high, and activity is strong, and consumption of organic solvent is less
The method that active polysaccharide is extracted from marine alga.
The present invention is directed to the problem mentioned in background technology, and the technical scheme taken is:
The method that active polysaccharide is extracted from marine alga, concretely comprises the following steps:
1)Degreasing, de- glycosides:Marine alga is cleaned, is crushed, sieving obtains marine algae powder, add acetone-petroleum ether mixed-liquor return to take off
Fat, marine algae powder:Acetone-petroleum ether mixeding liquid volume ratio is 1:10 ~ 15, acetone:Petroleum ether volume ratio is 1:1.2~1:1.6.
Grease solubility in acetone-petroleum ether mixed liquor is big in marine alga, and marine alga degreasing rate is high.Suction filtration, adds alcohol reflux in filter residue,
Concentration of alcohol is 85 ~ 90%, filter residue:Ethanol volume ratio is 1:12 ~ 16, suction filtration, filter residue drying.Glycoside and alkaloid in marine alga
Solubility is high in ethanol, and polysaccharide does not dissolve in ethanol, and polysaccharide loss rate is few during removal of impurities;
2)Enzymolysis:Distilled water, filter residue are added in the filter residue that step 1 is finally obtained:Distilled water volume ratio:1:25~28.Complex enzyme
Preparation is digested, and complex enzyme addition is 3 ~ 5%, and hydrolysis temperature is 52 ~ 56 DEG C, and pH is 2.7 ~ 3.4, and enzymolysis time is 2.2 ~ 2.7h.
Complex enzyme formulation composition and its weight portion are:10 ~ 20 parts of cellulases, 10 ~ 15 parts of pectases, 1 ~ 3 part of NaCl, 0.001 ~
0.002 part of potassium pyrophosphate, 0.5 ~ 1 part of magnesium sulfate, 0.004 ~ 0.006 part of APP and 0.2 ~ 0.8 part of calcium dihydrogen phosphate.It is above-mentioned
Complex enzyme formulation digests the efficiency high of cellulose and pectin, and digests fully, and the destructive rate to cell membrane is high.It is ultrasonic-wave assisted
Molten, supersonic frequency is 260 ~ 320W, and ultrasonic time is 20 ~ 30min.Enzymolysis terminates rear boiling water bath and goes out enzyme, and filtering takes filtrate.Enzyme
Cellulose and hydrolyzed pectin are destroyed cell wall structure by solution, and are aided with ultrasonic wave extraction, and active polysaccharide flows from alginic cell
Go out to be dissolved in the water, the recovery rate of active polysaccharide is high;
3)Deproteinized:Sevage reagents, Sevage reagents are added in filtrate:Filtrate volume ratio is 1:3 ~ 5, protein exists
Can be denatured in Sevage reagents, solubility declines in water, stirring removes lower floor's liquid and flocky precipitate, in supernatant liquid
Middle addition ethanol, concentration of alcohol is 90 ~ 98%, supernatant liquid:Ethanol volume ratio is 1:3.7~4.5.Stirring, suction filtration takes filter residue, first
With 35 ~ 40 times of absolute ethanol washing of filter residue volume, then washed with 30 ~ 32 times of anhydrous propanone, revolving is dried to obtain rough sea
Algae active polysaccharide powder;
4)Ultrafiltration:Distilled water, ultrafiltration are added in rough marine alga active polysaccharide powder, filter membrane specification is 3500 ~ 4000Da, and activity is more
Glycan molecule amount is more than 3500 ~ 4000Da, and ultrafiltration can remove the small molecular weight impurity for being dissolved in water.Non- filtered solution is taken, powder is dried to obtain;
5)Gel chromatography:Distilled water, powder are added in powder:Distilled water volume ratio is 1:0.02 ~ 0.05, inject DEAE- fine jades
Chromatographic isolation in sepharose post, first rinses 4.2 ~ 4.6h with 1.2 ~ 1.4mL/min of distilled water, then with 0.5MNaCl with same
Flow velocity continues to rinse 2.2 ~ 2.5h.Heterogeneity molecular size range is different, and gel chromatography is more by marine alga according to the size of molecular weight
Each composition is separated in sugar aqueous solution, is detected using benzene phenol-sulfuric acid method, and collection has the eluent of chromogenic reaction, to marine alga
Active polysaccharide is further isolated and purified;
6)Desalt, dry:Eluent is placed in bag filter and is dialysed, filter membrane specification takes non-filtered solution for 3500 ~ 4000Da, will be solidifying
The NaCl that glue-line is introduced when analysing is removed.It is concentrated under reduced pressure, it is concentrated under reduced pressure into the 1/100 ~ 1/200 of non-filtered solution volume, freeze-drying
Obtain active polysaccharide powder.The purity of the active polysaccharide powder for obtaining is very high, and activity is strong, organic solvent-free residual, green health,
It is adapted to be applied to cosmetics, medicine, field of food.The recyclable recycling of organic solvent used in preparation process, to environment
Pollution is small.
Compared with prior art, the advantage of the invention is that:With acetone-petroleum ether mixed-liquor return degreasing, returned with ethanol
Stream de- glycoside and alkaloid, carry out removal of impurities;Complex enzyme formulation digests the efficiency high of cellulose and pectin, and digests fully, right
The destructive rate of cell membrane is high;Cellulose and hydrolyzed pectin are destroyed cell wall structure by enzymolysis, and are aided with ultrasonic wave extraction, burnt
Potassium phosphate and APP produce unexpected effect by magnesium sulfate acetin, promote active polysaccharide to be flowed out from alginic cell
It is dissolved in the water, the recovery rate of active polysaccharide is high;Make protein denaturation with Sevage reagents, remove removing protein;Using ultrafiltration, coagulate
Glue-line is analysed and dialysis is isolated and purified to rough marine alga active polysaccharide powder, and the marine alga active polysaccharide purity for finally obtaining is very
Height, activity is strong, and organic solvent-free residual, green health is adapted to be applied to cosmetics, medicine, field of food;Used in preparation process
The recyclable recycling of organic solvent for arriving, the pollution to environment is small;With low cost, active polysaccharide economic worth is high, is adapted to factory
Metaplasia is produced.
Specific embodiment
The present invention program is described further below by embodiment:
Embodiment 1:
The method that active polysaccharide is extracted from marine alga, concretely comprises the following steps:
1)Degreasing, de- glycosides:Marine alga is cleaned, is crushed, sieving obtains marine algae powder, add acetone-petroleum ether mixed-liquor return to take off
Fat, marine algae powder:Acetone-petroleum ether mixeding liquid volume ratio is 1:10 ~ 15, acetone:Petroleum ether volume ratio is 1:1.2~1:1.6.
Grease solubility in acetone-petroleum ether mixed liquor is big in marine alga, and marine alga degreasing rate is high.Suction filtration, adds alcohol reflux in filter residue,
Concentration of alcohol is 85 ~ 90%, filter residue:Ethanol volume ratio is 1:12 ~ 16, suction filtration, filter residue drying.Glycoside and alkaloid in marine alga
Solubility is high in ethanol, and polysaccharide does not dissolve in ethanol, and polysaccharide loss rate is few during removal of impurities;
2)Enzymolysis:Distilled water, filter residue are added in the filter residue that step 1 is finally obtained:Distilled water volume ratio:1:25~28.Complex enzyme
Preparation is digested, and complex enzyme addition is 3 ~ 5%, and hydrolysis temperature is 52 ~ 56 DEG C, and pH is 2.7 ~ 3.4, and enzymolysis time is 2.2 ~ 2.7h.
Complex enzyme formulation composition and its weight portion are:10 ~ 20 parts of cellulases, 10 ~ 15 parts of pectases, 1 ~ 3 part of NaCl, 0.001 ~
0.002 part of potassium pyrophosphate, 0.5 ~ 1 part of magnesium sulfate, 0.004 ~ 0.006 part of APP and 0.2 ~ 0.8 part of calcium dihydrogen phosphate.It is above-mentioned
Complex enzyme formulation digests the efficiency high of cellulose and pectin, and digests fully, and the destructive rate to cell membrane is high.It is ultrasonic-wave assisted
Molten, supersonic frequency is 260 ~ 320W, and ultrasonic time is 20 ~ 30min.Enzymolysis terminates rear boiling water bath and goes out enzyme, and filtering takes filtrate.Enzyme
Cellulose and hydrolyzed pectin are destroyed cell wall structure by solution, and are aided with ultrasonic wave extraction, and active polysaccharide flows from alginic cell
Go out to be dissolved in the water, the recovery rate of active polysaccharide is high;
3)Deproteinized:Sevage reagents, Sevage reagents are added in filtrate:Filtrate volume ratio is 1:3 ~ 5, protein exists
Can be denatured in Sevage reagents, solubility declines in water, stirring removes lower floor's liquid and flocky precipitate, in supernatant liquid
Middle addition ethanol, concentration of alcohol is 90 ~ 98%, supernatant liquid:Ethanol volume ratio is 1:3.7~4.5.Stirring, suction filtration takes filter residue, first
With 35 ~ 40 times of absolute ethanol washing of filter residue volume, then washed with 30 ~ 32 times of anhydrous propanone, revolving is dried to obtain rough sea
Algae active polysaccharide powder;
4)Ultrafiltration:Distilled water, ultrafiltration are added in rough marine alga active polysaccharide powder, filter membrane specification is 3500 ~ 4000Da, and activity is more
Glycan molecule amount is more than 3500 ~ 4000Da, and ultrafiltration can remove the small molecular weight impurity for being dissolved in water.Non- filtered solution is taken, powder is dried to obtain;
5)Gel chromatography:Distilled water, powder are added in powder:Distilled water volume ratio is 1:0.02 ~ 0.05, inject DEAE- fine jades
Chromatographic isolation in sepharose post, first rinses 4.2 ~ 4.6h with 1.2 ~ 1.4mL/min of distilled water, then with 0.5MNaCl with same
Flow velocity continues to rinse 2.2 ~ 2.5h.Heterogeneity molecular size range is different, and gel chromatography is more by marine alga according to the size of molecular weight
Each composition is separated in sugar aqueous solution, is detected using benzene phenol-sulfuric acid method, and collection has the eluent of chromogenic reaction, to marine alga
Active polysaccharide is further isolated and purified;
6)Desalt, dry:Eluent is placed in bag filter and is dialysed, filter membrane specification takes non-filtered solution for 3500 ~ 4000Da, will be solidifying
The NaCl that glue-line is introduced when analysing is removed.It is concentrated under reduced pressure, it is concentrated under reduced pressure into the 1/100 ~ 1/200 of non-filtered solution volume, freeze-drying
Obtain active polysaccharide powder.The purity of the active polysaccharide powder for obtaining is very high, and activity is strong, organic solvent-free residual, green health,
It is adapted to be applied to cosmetics, medicine, field of food.The recyclable recycling of organic solvent used in preparation process, to environment
Pollution is small.
Embodiment 2:
The method that active polysaccharide is extracted from marine alga, most preferably step is:
1)Degreasing, de- glycosides:Marine alga is cleaned, is crushed, sieving obtains marine algae powder, add acetone-petroleum ether mixed-liquor return to take off
Fat, marine algae powder:Acetone-petroleum ether mixeding liquid volume ratio is 1:12, acetone:Petroleum ether volume ratio is 1:1.4.Grease in marine alga
Solubility is big in acetone-petroleum ether mixed liquor, and marine alga degreasing rate is high.Suction filtration, adds the alcohol reflux, concentration of alcohol to be in filter residue
88%, filter residue:Ethanol volume ratio is 1:15, suction filtration, filter residue drying.Solubility is high in ethanol for glycoside and alkaloid in marine alga,
Polysaccharide does not dissolve in ethanol, and polysaccharide loss rate is few during removal of impurities;
2)Enzymolysis:Distilled water, filter residue are added in the filter residue that step 1 is finally obtained:Distilled water volume ratio is 1:27.Complex enzyme system
Agent is digested, and complex enzyme addition is 4%, and hydrolysis temperature is 54 DEG C, and pH is 3.1, and enzymolysis time is 2.5h.Complex enzyme formulation composition
And its most preferably weight portion is:17 parts of cellulases, 12 parts of pectases, 2 parts of NaCl, 0.002 part of potassium pyrophosphate, 0.7 part of sulfuric acid
Magnesium, 0.005 part of APP and 0.5 part of calcium dihydrogen phosphate.The efficiency high of above-mentioned complex enzyme formulation enzymolysis cellulose and pectin, and
And enzymolysis is abundant, the destructive rate to cell membrane is high.Ultrasonic wave hydrotropy, supersonic frequency is 290W, and ultrasonic time is 24min.Enzymolysis
Boiling water bath goes out enzyme after end, filtering, takes filtrate.Cellulose and hydrolyzed pectin are destroyed cell wall structure, and be aided with super by enzymolysis
Sound wave is extracted, and active polysaccharide flows out from alginic cell and is dissolved in the water, and the recovery rate of active polysaccharide is high;
3)Deproteinized:Sevage reagents, Sevage reagents are added in filtrate:Filtrate volume ratio is 1:4, protein exists
Can be denatured in Sevage reagents, solubility declines in water, stirring removes lower floor's liquid and flocky precipitate, in supernatant liquid
Middle addition ethanol, concentration of alcohol is 95%, supernatant liquid:Ethanol volume ratio is 1:4.2.Stirring, suction filtration takes filter residue, first uses filter residue
The absolute ethanol washing that 37 times of volume, then washed with 31 times of anhydrous propanone, revolving is dried to obtain rough marine alga active polysaccharide powder
End;
4)Ultrafiltration:Distilled water, ultrafiltration are added in rough marine alga active polysaccharide powder, filter membrane specification is 3500Da, active polysaccharide point
Son amount is more than 3500Da, and ultrafiltration can remove the small molecular weight impurity for being dissolved in water.Non- filtered solution is taken, powder is dried to obtain;
5)Gel chromatography:Distilled water, powder are added in powder:Distilled water volume ratio is 1:0.03, injection DEAE- agarose coagulate
Chromatographic isolation in glue post, first rinses 4.4h with distilled water 1.3mL/min, then is continued to rinse with same flow velocity with 0.5MNaCl
2.4h.Heterogeneity molecular size range is different, and gel chromatography is according to the size of molecular weight by each composition in the algal polysaccharides aqueous solution
Separate, detected using benzene phenol-sulfuric acid method, collection has the eluent of chromogenic reaction, and marine alga active polysaccharide is further divided
From purifying;
6)Desalt, dry:Eluent is placed in bag filter and is dialysed, filter membrane specification non-filtered solution for 3500Da takes, by gel layer
The NaCl introduced during analysis is removed.It is concentrated under reduced pressure, the 1/140 of non-filtered solution volume is concentrated under reduced pressure into, it is many that freeze-drying obtains activity
Icing Sugar end.The purity of the active polysaccharide powder for obtaining is very high, and activity is strong, and organic solvent-free residual, green health is adapted to be applied to
Cosmetics, medicine, field of food.The recyclable recycling of organic solvent used in preparation process, the pollution to environment is small.
Routine operation in operating procedure of the invention is well known to those skilled in the art, and is not repeated herein.
Embodiment described above has been described in detail to technical scheme, it should be understood that the above is only
It is specific embodiment of the invention, is not intended to limit the invention, all any modifications made in spirit of the invention,
Supplement or similar fashion replacement etc., should be included within the scope of the present invention.
Claims (9)
1. the method that active polysaccharide is extracted from marine alga, it is characterised in that following steps:
1)Degreasing, de- glycosides:Marine alga is cleaned, is crushed, sieving obtains marine algae powder, add acetone-petroleum ether mixed-liquor return to take off
Fat, suction filtration adds alcohol reflux, suction filtration, filter residue drying in filter residue;
2)Enzymolysis:Distilled water and complex enzyme formulation enzymolysis, ultrasonic wave hydrotropy, enzymolysis are added in the filter residue that step 1 is finally obtained
Go out enzyme after end, filtering, takes filtrate;
3)Deproteinized:Sevage reagents are added in filtrate, stirring removes lower floor's liquid and flocky precipitate, in supernatant liquid
Middle addition ethanol, stirring, suction filtration takes filter residue, is washed with absolute ethyl alcohol and anhydrous propanone, and revolving is dried to obtain rough marine alga activity
Polysaccharide powder;
4)Ultrafiltration:Distilled water is added in rough marine alga active polysaccharide powder, ultrafiltration takes non-filtered solution, is dried to obtain powder;
5)Gel chromatography:Distilled water is added in powder, gel chromatography, benzene phenol-sulfuric acid method detection, collection has washing for chromogenic reaction
De- liquid;
6)Desalt, dry:Eluent is placed in bag filter and is dialysed, take non-filtered solution, be concentrated under reduced pressure, freeze-drying obtains activity
Polysaccharide powder.
2. it is according to claim 1 from marine alga extract active polysaccharide method, it is characterised in that:In described step 1
Marine algae powder:Acetone-petroleum ether mixeding liquid volume ratio is 1:10 ~ 15, acetone:Petroleum ether volume ratio is 1:1.2~1:1.6, ethanol
Concentration is 85 ~ 90%, filter residue:Ethanol volume ratio is 1:12~16.
3. it is according to claim 1 from marine alga extract active polysaccharide method, it is characterised in that:In described step 2
Filter residue:Distilled water volume ratio:1:25 ~ 28, complex enzyme addition is 3 ~ 5%, and hydrolysis temperature is 52 ~ 56 DEG C, and pH is 2.7 ~ 3.4, enzyme
The solution time is 2.2 ~ 2.7h.
4. it is according to claim 1 from marine alga extract active polysaccharide method, it is characterised in that:In described step 2
Complex enzyme formulation composition and its weight portion are:10 ~ 20 parts of cellulases, 10 ~ 15 parts of pectases, 1 ~ 3 part of NaCl, 0.001 ~
0.002 part of potassium pyrophosphate, 0.5 ~ 1 part of magnesium sulfate, 0.004 ~ 0.006 part of APP and 0.2 ~ 0.8 part of calcium dihydrogen phosphate.
5. it is according to claim 1 from marine alga extract active polysaccharide method, it is characterised in that:In described step 3
Sevage reagents:Filtrate volume ratio is 1:3 ~ 5, concentration of alcohol is 90 ~ 98%, supernatant liquid:Ethanol volume ratio is 1:3.7~
4.5。
6. it is according to claim 1 from marine alga extract active polysaccharide method, it is characterised in that:In described step 3
First with the absolute ethanol washing of 35 ~ 40 times of filter residue volume, then washed with 30 ~ 32 times of anhydrous propanone.
7. it is according to claim 1 from marine alga extract active polysaccharide method, it is characterised in that:Described step 4 with
Filter membrane specification is 3500 ~ 4000Da in step 6.
8. it is according to claim 1 from marine alga extract active polysaccharide method, it is characterised in that:In described step 5
Powder:Distilled water volume ratio is 1:0.02 ~ 0.05, chromatographic isolation in injection DEAE- agarose Gel columns first uses distilled water
1.2 ~ 1.4mL/min rinses 4.2 ~ 4.6h, then is continued to rinse 2.2 ~ 2.5h with same flow velocity with 0.5MNaCl.
9. it is according to claim 1 from marine alga extract active polysaccharide method, it is characterised in that:In described step 6
It is concentrated under reduced pressure into the 1/100 ~ 1/200 of non-filtered solution volume.
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CN108746658A (en) * | 2018-05-24 | 2018-11-06 | 西安理工大学 | A kind of preparation method of the algal polysaccharides nanogold particle with antibacterial activity |
CN108997511A (en) * | 2018-09-07 | 2018-12-14 | 南阳理工学院 | One main laminaria active polysaccharide and preparation method thereof |
CN109734820A (en) * | 2018-11-22 | 2019-05-10 | 浙江工业大学 | A kind of preparation method of small molecule fucoidin |
CN109608559A (en) * | 2019-01-15 | 2019-04-12 | 广东海洋大学 | A method of extracting active polysaccharide from seaweed |
CN111318050A (en) * | 2020-03-05 | 2020-06-23 | 广西科学院 | Method for comprehensively extracting polyunsaturated fatty acid and polysaccharide from seaweed |
CN111318050B (en) * | 2020-03-05 | 2022-07-01 | 广西科学院 | Method for comprehensively extracting polyunsaturated fatty acid and polysaccharide from seaweed |
CN115281194A (en) * | 2022-08-26 | 2022-11-04 | 山东康惠植物保护有限公司 | Bactericide containing fluxapyroxad and trifloxystrobin |
CN115281194B (en) * | 2022-08-26 | 2023-09-19 | 山东康惠植物保护有限公司 | Bactericide containing fluxapyroxad and trifloxystrobin |
CN117247849A (en) * | 2023-11-20 | 2023-12-19 | 山东悦翔生物有限公司 | Microalgae extraction method |
CN117247849B (en) * | 2023-11-20 | 2024-02-06 | 山东悦翔生物有限公司 | Microalgae extraction method |
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