CN105907831A - Preparation method of high-purity agaricus blazei murrill polysaccharide - Google Patents

Preparation method of high-purity agaricus blazei murrill polysaccharide Download PDF

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CN105907831A
CN105907831A CN201610484563.1A CN201610484563A CN105907831A CN 105907831 A CN105907831 A CN 105907831A CN 201610484563 A CN201610484563 A CN 201610484563A CN 105907831 A CN105907831 A CN 105907831A
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agaricus blazei
blazei murrill
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郭迎庆
张明
孟浩影
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    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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Abstract

The invention discloses a preparation method of high-purity agaricus blazei murrill polysaccharide, belonging to the technical field of polysaccharide preparation. The preparation method comprises the steps of: pulverizing agaricus blazei murrill, carrying out pretreatment with sodium hydroxide, adding a mixed enzyme solution, disinfecting the reaction solution and inactivating enzymes after reaction; centrifuging the reaction solution to obtain an extracting solution of the agaricus blazei murrill; adding polyacrylamide, ferrous sulfate and the like for impurity removal and color removal; carrying out extraction in a carbon dioxide supercritical extractor to obtain a mixed solution; adding maleic acid for reaction, filtering the mixed solution to obtain filtrate, adding tetraethylammonium hydroxide for reaction; cooling the solution with liquid nitrogen, and carrying out filtration, reduced pressure distillation and drying to obtain the high-purity agaricus blazei murrill polysaccharide. Proven by examples, the preparation method has the advantages of simple operation and mild reaction conditions, the whole preparation process is free of addition of any organic solvents, so that the yield of the finally prepared agaricus blazei murrill polysaccharide reaches over 90%, the purity is not lower than 95%, and the preparation method lays a foundation for industrialized development of the agaricus blazei murrill polysaccharide in the field of medicine and the health product industry.

Description

A kind of preparation method of high-purity Agaricus Blazei Murrill polysaccharide
Technical field
The invention discloses the preparation method of a kind of high-purity Agaricus Blazei Murrill polysaccharide, belong to polysaccharide preparing technical field.
Background technology
Agaricus blazei Murrill has another name called Brazilian mushroom, is a kind of China dietotherapeutic leaved for development bacterium, and it originates in the ground such as Brazil of North American southern, Peru.Account for 40%～45% containing protein in every 100 grams of dry mushrooms of Agaricus blazei Murrill, saccharide accounts for 38%～45%, crude fat accounts for 3%～4%, crude fibre 6.73% and abundant vitamin thereof and trace element, enriches as food nutrition.Agaricus blazei Murrill contains abundant bioactive substance, can use as medicine and health product；Primary bioactivity material includes polysaccharide, sterols, nucleic acid, unsaturated fatty acid, dietary fiber etc., and according to Japan Report, the antitumaous effect of Agaricus blazei Murrill is substantially better than other the 14 kinds macro fungis having antitumaous effect, occupies the anticancer first place of fungus.Therefore, medical value is by the very big concern of cuisines, medical science and pharmacy circle.
Agaricus Blazei Murrill polysaccharide is the effective active composition extracted from Agaricus blazei Murrill sporophore, its fragrant odour, effect is notable, have and body is had the effect strengthening energy, and the growth of tumor cell can be suppressed, there is antitumor action, in the case of those cannot be carried out performing the operation and also cannot use radiation or chemotherapy, it is effective too, and Agaricus blazei Murrill and extract thereof can also the most effectively alleviate the side effect that the traditional therapies such as chemotherapy are brought.It addition, also there is blood sugar lowering, improve diabetes, cholesterol reducing, delay the effect of arteriosclerosis.
Agaricus Blazei Murrill polysaccharide is in addition to can be as the healing potion in terms of antiviral property disease, parasiticide, anti-repulsion and anti-curing oncoma, it is alternatively arranged as immunostimulant and improves humoral immunity level, its extract can remarkably promote interleukin-6 (IL-6) and interferon-γ (IFN-γ) level, reduce in vitro and in vivo IL-4 level, thus immunologic function is adjusted.It is noted that Agaricus Blazei Murrill polysaccharide can be obviously enhanced the effect of vaccine as immunological adjuvant, there is the most wide application prospect.
At present, on domestic and international market, Agaricus Blazei Murrill polysaccharide is mainly content 10～the specification of 50%, extracting method many employings Hot water extraction extracts (subject matter be extraction efficiency is low, poor stability), and polysaccharide purification method generally uses ethanol precipitation, Sevgag method, major defect to be that organic solvent introduces and the problem of residual.
Summary of the invention
The technical problem that present invention mainly solves: for current traditional method during preparing Agaricus Blazei Murrill polysaccharide, Hot water extraction extraction efficiency is low, poor stability, and ethanol precipitation, Sevgag method use substantial amounts of organic solvent, make the present situation that solvent residual amount in product is bigger, it is provided that the preparation method of a kind of high-purity Agaricus Blazei Murrill polysaccharide.Agaricus blazei Murrill is pulverized by the method, after carrying out pretreatment with sodium hydroxide, adds mixed enzyme solution, by reactant liquor sterilization enzyme inactivation after reaction, centrifugal to obtain Agaricus blazei Murrill extracting solution, to add the remove impurity such as polyacrylamide, ferrous sulfate except color, then to be placed in carbon dioxide supercritical fluid extraction instrument and extract, obtain mixed solution, add maleic acid reaction, be filtrated to get filtrate, add tetraethyl ammonium hydroxide reaction, and make solution lower the temperature with liquid nitrogen, through filtering, reduce pressure distillation, being dried to obtain high-purity Agaricus Blazei Murrill polysaccharide.The present invention is easy and simple to handle, reaction condition is gentle, without the addition of any organic solvent in whole preparation process so that the final Agaricus Blazei Murrill polysaccharide yield prepared has reached more than 90%, purity is not less than 95%, lays a good foundation for Agaricus Blazei Murrill polysaccharide industrialized developing in field of medicaments and health products trade.
In order to solve above-mentioned technical problem, the technical solution adopted in the present invention is:
(1) 10～15g sucrose, 2～4g sodium nitrate, 1～3gMgSO are weighed successively4·7H2O, 1～3gKCl, 0.8～1.0gFeSO4·7H2O, 10～20g agar joins in culture dish, adds 800～1000mL deionized waters, stirs 10～20min, makes culture medium, cultivates strain；
(2) successively inoculation 2～4 strain Trichoderma spp., 2～4 strain aspergillus nigers, 2～4 bacillus in culture medium, at 38～40 DEG C, by shaking table shaken cultivation 12～14h, culture fluid is centrifuged under 7000～9000r/min 30～40min, abandoning supernatant, obtains mixed bacteria；
(3) pipette 10～20% above-mentioned mixed bacteria join in fermentation tank, sequentially adding 5～10g glucoses, 1～3g Carnis Bovis seu Bubali cream, 200～300mLpH is 6.8～7.0 phosphate buffered solution, stirring 10～20min, at 50～60 DEG C, by shaking table shaken cultivation 48～50h, culture fluid is centrifuged under 6000～8000r/min 20～30min, discards lower floor's impurity, obtain mixed enzyme solution；
(4) weigh 10～20g dry Agaricus blazei Murrill, pulverize with pulverizer, then Agaricus blazei Murrill powder is joined in beaker, being placed in water-bath by beaker, control temperature, at 45～55 DEG C, adds 200～300mL0.01mol/L sodium hydroxide solutions in beaker, stirring 1～2h, add above-mentioned mixed enzyme solution, stirring reaction 3～4h, temperature is increased to 90～100 DEG C, stirring 10～20min, filter, obtain filtrate, i.e. Agaricus blazei Murrill extracting solution；
(5) in above-mentioned Agaricus blazei Murrill extracting solution, 1～3g polyacrylamide, 1～3g ferrous sulfate, 1～3g activated carbon it are sequentially added into, stirring 10～20min, filter, obtain filtrate, filtrate is placed in carbon dioxide supercritical fluid extraction instrument, controlling extracting pressure is 20～30MPa, and extraction temperature is 45～55 DEG C, CO2Flow be 25～35kg/h, extract 20～30min, obtain mixed solution；
(6) above-mentioned mixed solution is joined in beaker, beaker is placed in ice-water bath, control temperature at 0～5 DEG C, 1～3g maleic acid is added in above-mentioned mixed solution, stirring reaction 1～2h, filter, obtain filtrate, 2～4g tetraethyl ammonium hydroxides are added in filtrate, stirring reaction 1～2h, by on 0.1～0.3L liquid nitrogen spraying to beaker, beaker temperature is made to be down to-30～-20 DEG C, stirring 10～20min, filter, obtain filtrate, i.e. Agaricus Blazei Murrill polysaccharide solution, Agaricus Blazei Murrill polysaccharide solution is carried out decompression distillation and obtains white solid, it is dried 2～3h in white solid is placed in 95～105 DEG C of baking ovens, obtain white powder, i.e. a kind of high-purity Agaricus Blazei Murrill polysaccharide.
The application process of the present invention is: the high-purity Agaricus Blazei Murrill polysaccharide present invention prepared makes Agaricus Blazei Murrill polysaccharide capsule by medical capsule machine processed, in the most each capsule, in Agaricus Blazei Murrill polysaccharide, content is 0.1～0.15g, time edible, it be equipped with warm water and be administered orally, it is proposed that every day is the most edible, the most edible 1～3, the most edible 7～14 days, immune function of human body can be adjusted, especially cancer patient, anticancer effect is the most notable, it is possible to decrease more than 75% cancer return rate.
The invention has the beneficial effects as follows:
(1) present invention is easy and simple to handle, and reaction condition is gentle, without the addition of any organic solvent in whole preparation process so that the final Agaricus Blazei Murrill polysaccharide yield prepared has reached more than 90%, and purity is not less than 95%；
(2) the Agaricus Blazei Murrill polysaccharide requisite quality that the present invention prepares, lays a good foundation for Agaricus Blazei Murrill polysaccharide industrialized developing in field of medicaments and health products trade.
Detailed description of the invention
Weigh 10～15g sucrose, 2～4g sodium nitrate, 1～3gMgSO the most successively4·7H2O, 1～3gKCl, 0.8～1.0gFeSO4·7H2O, 10～20g agar joins in culture dish, adds 800～1000mL deionized waters, stirs 10～20min, makes culture medium, cultivates strain；The most successively inoculation 2～4 strain Trichoderma spp., 2～4 strain aspergillus nigers, 2～4 bacillus in culture medium, at 38～40 DEG C, by shaking table shaken cultivation 12～14h, culture fluid is centrifuged under 7000～9000r/min 30～40min, abandoning supernatant, obtains mixed bacteria；Next pipette 10～20% above-mentioned mixed bacteria join in fermentation tank, sequentially adding 5～10g glucoses, 1～3g Carnis Bovis seu Bubali cream, 200～300mLpH is 6.8～7.0 phosphate buffered solution, stirring 10～20min, at 50～60 DEG C, by shaking table shaken cultivation 48～50h, culture fluid is centrifuged under 6000～8000r/min 20～30min, discards lower floor's impurity, obtain mixed enzyme solution；Then weigh 10～20g dry Agaricus blazei Murrill, pulverize with pulverizer, then Agaricus blazei Murrill powder is joined in beaker, being placed in water-bath by beaker, control temperature, at 45～55 DEG C, adds 200～300mL0.01mol/L sodium hydroxide solutions in beaker, stirring 1～2h, add above-mentioned mixed enzyme solution, stirring reaction 3～4h, temperature is increased to 90～100 DEG C, stirring 10～20min, filter, obtain filtrate, i.e. Agaricus blazei Murrill extracting solution；1～3g polyacrylamide, 1～3g ferrous sulfate, 1～3g activated carbon it is sequentially added into afterwards in above-mentioned Agaricus blazei Murrill extracting solution, stirring 10～20min, filter, obtain filtrate, filtrate is placed in carbon dioxide supercritical fluid extraction instrument, controlling extracting pressure is 20～30MPa, and extraction temperature is 45～55 DEG C, CO2Flow be 25～35kg/h, extract 20～30min, obtain mixed solution；Finally above-mentioned mixed solution is joined in beaker, beaker is placed in ice-water bath, control temperature at 0～5 DEG C, 1～3g maleic acid is added in above-mentioned mixed solution, stirring reaction 1～2h, filter, obtain filtrate, 2～4g tetraethyl ammonium hydroxides are added in filtrate, stirring reaction 1～2h, by on 0.1～0.3L liquid nitrogen spraying to beaker, beaker temperature is made to be down to-30～-20 DEG C, stirring 10～20min, filter, obtain filtrate, i.e. Agaricus Blazei Murrill polysaccharide solution, Agaricus Blazei Murrill polysaccharide solution is carried out decompression distillation and obtains white solid, it is dried 2～3h in white solid is placed in 95～105 DEG C of baking ovens, obtain white powder, i.e. a kind of high-purity Agaricus Blazei Murrill polysaccharide.
Example 1
Weigh 10g sucrose, 2g sodium nitrate, 1gMgSO the most successively4·7H2O、1gKCl、0.8gFeSO4·7H2O, 10g agar joins in culture dish, adds 800mL deionized water, stirs 10min, makes culture medium, cultivates strain；Inoculate on 2 strain Trichoderma spp., 2 strain aspergillus nigers, 2 bacillus to culture medium the most successively, at 38 DEG C, by shaking table shaken cultivation 12h, by culture fluid centrifugal 30min, abandoning supernatant under 7000r/min, obtain mixed bacteria；Next pipette 10% above-mentioned mixed bacteria and join in fermentation tank, sequentially add 5g glucose, 1g Carnis Bovis seu Bubali cream, 200mLpH are 6.8 phosphate buffered solution, stirring 10～20min, at 50 DEG C, by shaking table shaken cultivation 48h, by culture fluid centrifugal 20min under 6000r/min, discard lower floor's impurity, obtain mixed enzyme solution；Then weigh the dry Agaricus blazei Murrill of 10g, pulverize with pulverizer, then Agaricus blazei Murrill powder is joined in beaker, being placed in water-bath by beaker, control temperature, at 45 DEG C, adds 200mL0.01mol/L sodium hydroxide solution in beaker, stirring 1h, add above-mentioned mixed enzyme solution, stirring reaction 3h, temperature is increased to 90 DEG C, stirring 10min, filter, obtain filtrate, i.e. Agaricus blazei Murrill extracting solution；In above-mentioned Agaricus blazei Murrill extracting solution, it is sequentially added into 1g polyacrylamide, 1g ferrous sulfate, 1g activated carbon afterwards, stirs 10min, filter, obtaining filtrate, filtrate be placed in carbon dioxide supercritical fluid extraction instrument, control extracting pressure is 20MPa, extraction temperature is 45 DEG C, CO2Flow be 25kg/h, extract 20min, obtain mixed solution；Finally above-mentioned mixed solution is joined in beaker, beaker is placed in ice-water bath, control temperature at 0 DEG C, 1g maleic acid is added in above-mentioned mixed solution, stirring reaction 1h, filter, obtain filtrate, 2g tetraethyl ammonium hydroxide is added in filtrate, stirring reaction 1h, by on 0.1L liquid nitrogen spraying to beaker, beaker temperature is made to be down to-30 DEG C, stirring 10min, filter, obtain filtrate, i.e. Agaricus Blazei Murrill polysaccharide solution, Agaricus Blazei Murrill polysaccharide solution is carried out decompression distillation and obtains white solid, it is dried 2h in white solid is placed in 95 DEG C of baking ovens, obtain white powder, i.e. a kind of high-purity Agaricus Blazei Murrill polysaccharide.
This example operation is easy, and during use, the high-purity Agaricus Blazei Murrill polysaccharide present invention prepared makes Agaricus Blazei Murrill polysaccharide capsule by medical capsule machine processed, in the most each capsule, in Agaricus Blazei Murrill polysaccharide, content is 0.1g, time edible, it be equipped with warm water and be administered orally, it is proposed that every day is the most edible, the most edible 1, the most edible 7 days, immune function of human body can be adjusted, especially cancer patient, anticancer effect is the most notable, it is possible to decrease 75% cancer return rate.
Example 2
Weigh 13g sucrose, 3g sodium nitrate, 2gMgSO the most successively4·7H2O、2gKCl、0.9gFeSO4·7H2O, 15g agar joins in culture dish, adds 900mL deionized water, stirs 15min, makes culture medium, cultivates strain；Inoculate on 3 strain Trichoderma spp., 3 strain aspergillus nigers, 3 bacillus to culture medium the most successively, at 39 DEG C, by shaking table shaken cultivation 13h, by culture fluid centrifugal 35min, abandoning supernatant under 8000r/min, obtain mixed bacteria；Next pipette 15% above-mentioned mixed bacteria and join in fermentation tank, sequentially add 8g glucose, 2g Carnis Bovis seu Bubali cream, 250mLpH are 6.9 phosphate buffered solution, stirring 15min, at 55 DEG C, by shaking table shaken cultivation 49h, by culture fluid centrifugal 25min under 7000r/min, discard lower floor's impurity, obtain mixed enzyme solution；Then weigh the dry Agaricus blazei Murrill of 15g, pulverize with pulverizer, then Agaricus blazei Murrill powder is joined in beaker, being placed in water-bath by beaker, control temperature, at 50 DEG C, adds 250mL0.01mol/L sodium hydroxide solution in beaker, stirring 1.5h, add above-mentioned mixed enzyme solution, stirring reaction 3.5h, temperature is increased to 95 DEG C, stirring 15min, filter, obtain filtrate, i.e. Agaricus blazei Murrill extracting solution；In above-mentioned Agaricus blazei Murrill extracting solution, it is sequentially added into 2g polyacrylamide, 2g ferrous sulfate, 2g activated carbon afterwards, stirs 15min, filter, obtaining filtrate, filtrate be placed in carbon dioxide supercritical fluid extraction instrument, control extracting pressure is 25MPa, extraction temperature is 50 DEG C, CO2Flow be 30kg/h, extract 25min, obtain mixed solution；Finally above-mentioned mixed solution is joined in beaker, beaker is placed in ice-water bath, control temperature at 3 DEG C, 2g maleic acid is added in above-mentioned mixed solution, stirring reaction 1.5h, filter, obtain filtrate, 3g tetraethyl ammonium hydroxide is added in filtrate, stirring reaction 1.5h, by on 0.2L liquid nitrogen spraying to beaker, beaker temperature is made to be down to-25 DEG C, stirring 15min, filter, obtain filtrate, i.e. Agaricus Blazei Murrill polysaccharide solution, Agaricus Blazei Murrill polysaccharide solution is carried out decompression distillation and obtains white solid, it is dried 2.5h in white solid is placed in 100 DEG C of baking ovens, obtain white powder, i.e. a kind of high-purity Agaricus Blazei Murrill polysaccharide.
This example operation is easy, and during use, the high-purity Agaricus Blazei Murrill polysaccharide present invention prepared makes Agaricus Blazei Murrill polysaccharide capsule by medical capsule machine processed, in the most each capsule, in Agaricus Blazei Murrill polysaccharide, content is 0.13g, time edible, it be equipped with warm water and be administered orally, it is proposed that every day is the most edible, the most edible 2, the most edible 10 days, immune function of human body can be adjusted, especially cancer patient, anticancer effect is the most notable, it is possible to decrease 78% cancer return rate.
Example 3
Weigh 15g sucrose, 4g sodium nitrate, 3gMgSO the most successively4·7H2O、3gKCl、1.0gFeSO4·7H2O, 20g agar joins in culture dish, adds 1000mL deionized water, stirs 20min, makes culture medium, cultivates strain；Inoculate on 4 strain Trichoderma spp., 4 strain aspergillus nigers, 4 bacillus to culture medium the most successively, at 40 DEG C, by shaking table shaken cultivation 14h, by culture fluid centrifugal 40min, abandoning supernatant under 9000r/min, obtain mixed bacteria；Next pipette 20% above-mentioned mixed bacteria and join in fermentation tank, sequentially add 10g glucose, 3g Carnis Bovis seu Bubali cream, 300mLpH are 7.0 phosphate buffered solution, stirring 20min, at 60 DEG C, by shaking table shaken cultivation 50h, by culture fluid centrifugal 30min under 8000r/min, discard lower floor's impurity, obtain mixed enzyme solution；Then weigh the dry Agaricus blazei Murrill of 20g, pulverize with pulverizer, then Agaricus blazei Murrill powder is joined in beaker, being placed in water-bath by beaker, control temperature, at 55 DEG C, adds 300mL0.01mol/L sodium hydroxide solution in beaker, stirring 2h, add above-mentioned mixed enzyme solution, stirring reaction 4h, temperature is increased to 100 DEG C, stirring 20min, filter, obtain filtrate, i.e. Agaricus blazei Murrill extracting solution；In above-mentioned Agaricus blazei Murrill extracting solution, it is sequentially added into 3g polyacrylamide, 3g ferrous sulfate, 3g activated carbon afterwards, stirs 20min, filter, obtaining filtrate, filtrate be placed in carbon dioxide supercritical fluid extraction instrument, control extracting pressure is 30MPa, extraction temperature is 55 DEG C, CO2Flow be 35kg/h, extract 30min, obtain mixed solution；Finally above-mentioned mixed solution is joined in beaker, beaker is placed in ice-water bath, control temperature at 5 DEG C, 3g maleic acid is added in above-mentioned mixed solution, stirring reaction 2h, filter, obtain filtrate, 4g tetraethyl ammonium hydroxide is added in filtrate, stirring reaction 2h, by on 0.3L liquid nitrogen spraying to beaker, beaker temperature is made to be down to-20 DEG C, stirring 20min, filter, obtain filtrate, i.e. Agaricus Blazei Murrill polysaccharide solution, Agaricus Blazei Murrill polysaccharide solution is carried out decompression distillation and obtains white solid, it is dried 3h in white solid is placed in 105 DEG C of baking ovens, obtain white powder, i.e. a kind of high-purity Agaricus Blazei Murrill polysaccharide.
This example operation is easy, and during use, the high-purity Agaricus Blazei Murrill polysaccharide present invention prepared makes Agaricus Blazei Murrill polysaccharide capsule by medical capsule machine processed, in the most each capsule, in Agaricus Blazei Murrill polysaccharide, content is 0.15g, time edible, it be equipped with warm water and be administered orally, it is proposed that every day is the most edible, the most edible 3, the most edible 14 days, immune function of human body can be adjusted, especially cancer patient, anticancer effect is the most notable, it is possible to decrease 80% cancer return rate.

Claims (1)

1. the preparation method of a high-purity Agaricus Blazei Murrill polysaccharide, it is characterised in that concrete preparation process is:
(1) 10～15g sucrose, 2～4g sodium nitrate, 1～3gMgSO are weighed successively4·7H2O, 1～3gKCl, 0.8～1.0gFeSO4·7H2O, 10～20g agar joins in culture dish, adds 800～1000mL deionized waters, stirs 10～20min, makes culture medium, cultivates strain；
(2) successively inoculation 2～4 strain Trichoderma spp., 2～4 strain aspergillus nigers, 2～4 bacillus in culture medium, at 38～40 DEG C, by shaking table shaken cultivation 12～14h, culture fluid is centrifuged under 7000～9000r/min 30～40min, abandoning supernatant, obtains mixed bacteria；
(3) pipette 10～20% above-mentioned mixed bacteria join in fermentation tank, sequentially adding 5～10g glucoses, 1～3g Carnis Bovis seu Bubali cream, 200～300mLpH is 6.8～7.0 phosphate buffered solution, stirring 10～20min, at 50～60 DEG C, by shaking table shaken cultivation 48～50h, culture fluid is centrifuged under 6000～8000r/min 20～30min, discards lower floor's impurity, obtain mixed enzyme solution；
(4) weigh 10～20g dry Agaricus blazei Murrill, pulverize with pulverizer, then Agaricus blazei Murrill powder is joined in beaker, being placed in water-bath by beaker, control temperature, at 45～55 DEG C, adds 200～300mL0.01mol/L sodium hydroxide solutions in beaker, stirring 1～2h, add above-mentioned mixed enzyme solution, stirring reaction 3～4h, temperature is increased to 90～100 DEG C, stirring 10～20min, filter, obtain filtrate, i.e. Agaricus blazei Murrill extracting solution；
(5) in above-mentioned Agaricus blazei Murrill extracting solution, 1～3g polyacrylamide, 1～3g ferrous sulfate, 1～3g activated carbon it are sequentially added into, stirring 10～20min, filter, obtain filtrate, filtrate is placed in carbon dioxide supercritical fluid extraction instrument, controlling extracting pressure is 20～30MPa, and extraction temperature is 45～55 DEG C, CO2Flow be 25～35kg/h, extract 20～30min, obtain mixed solution；
(6) above-mentioned mixed solution is joined in beaker, beaker is placed in ice-water bath, control temperature at 0～5 DEG C, 1～3g maleic acid is added in above-mentioned mixed solution, stirring reaction 1～2h, filter, obtain filtrate, 2～4g tetraethyl ammonium hydroxides are added in filtrate, stirring reaction 1～2h, by on 0.1～0.3L liquid nitrogen spraying to beaker, beaker temperature is made to be down to-30～-20 DEG C, stirring 10～20min, filter, obtain filtrate, i.e. Agaricus Blazei Murrill polysaccharide solution, Agaricus Blazei Murrill polysaccharide solution is carried out decompression distillation and obtains white solid, it is dried 2～3h in white solid is placed in 95～105 DEG C of baking ovens, obtain white powder, i.e. a kind of high-purity Agaricus Blazei Murrill polysaccharide.
CN201610484563.1A 2016-06-28 2016-06-28 Preparation method of high-purity agaricus blazei murrill polysaccharide Withdrawn CN105907831A (en)

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CN106821102A (en) * 2016-12-29 2017-06-13 安徽比伦生活用纸有限公司 A kind of antibacterial facial tissue
CN112079939A (en) * 2020-08-25 2020-12-15 福州康来生物科技有限公司 Preparation method of agaricus blazei polysaccharide extract
CN112321643A (en) * 2020-10-20 2021-02-05 广药白云山化学制药(珠海)有限公司 Post-treatment method of Propofol tenofovir solution and preparation method of Propofol tenofovir
CN115364138A (en) * 2022-09-19 2022-11-22 博斯创想(上海)医疗科技有限公司 Compound bacterial liquid for improving chemotherapy side effects and preparation method thereof

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106821102A (en) * 2016-12-29 2017-06-13 安徽比伦生活用纸有限公司 A kind of antibacterial facial tissue
CN112079939A (en) * 2020-08-25 2020-12-15 福州康来生物科技有限公司 Preparation method of agaricus blazei polysaccharide extract
CN112321643A (en) * 2020-10-20 2021-02-05 广药白云山化学制药(珠海)有限公司 Post-treatment method of Propofol tenofovir solution and preparation method of Propofol tenofovir
CN112321643B (en) * 2020-10-20 2023-06-02 广药白云山化学制药(珠海)有限公司 Post-treatment method of propiophenone tenofovir solution and preparation method of propiophenone tenofovir
CN115364138A (en) * 2022-09-19 2022-11-22 博斯创想(上海)医疗科技有限公司 Compound bacterial liquid for improving chemotherapy side effects and preparation method thereof

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