CN104844721B - Extraction and separation method of Agrocybe aegirit polysaccharides - Google Patents
Extraction and separation method of Agrocybe aegirit polysaccharides Download PDFInfo
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Abstract
The invention belongs to the field of extraction and separation of polysaccharides, and discloses an extraction and separation method of Agrocybe aegirit polysaccharides. The extraction and separation method of Agrocybe aegirit polysaccharides comprises the following steps: 1, cutting fresh Agrocybe aegirit to form slices, pre-cooling the slices, and freeze-drying after the Agrocybe aegirit is frozen; 2, rapidly crushing the freeze-dried Agrocybe aegirit slices, adding an ethanol-water solution into the obtained Agrocybe aegirit powder, carrying out ultrasonic stirring extraction, centrifuging the obtained extraction liquid, and taking the obtained supernatant; 3, concentrating the supernatant until the weight of the obtained Agrocybe aegirit concentrate liquid is 2 times the weight of Agrocybe aegirit; 4, carrying out membrane filtration on the Agrocybe aegirit concentrate liquid through using a membrane with the molecular amount of 1000, and collecting the obtained trapped liquid; 5, allowing the trapped liquid to go through a membrane with the molecular amount of 200, and collecting the obtained new trapped liquid; 6, concentrating until no solvent exists; and 7, carrying out crystallizing purification on the obtained concentrate by using ethanol, filtering, and naturally drying to obtain the Agrocybe aegirit polysaccharides.
Description
Technical field
The invention belongs to extraction separation field, it is related to a kind of extraction separation method of polysaccharide, especially relates to a kind of Camellia sinensis
The extraction separation method of mushroom polysaccharide.
Background technology
Polysaccharide (polysaccharide) is to form glycosidic bond by dehydration between monosaccharide, and linearly or branch connects with glycosidic bond
The chain polymer connecing.Typically the sugar chain more than 20 glycosyls is then referred to as polysaccharide.In recent years, carbohydrate structure measures and raw
Thing activity research achieves obvious progress, and the true saccharide that discloses of many experiments is important information molecule, participate in many physiology and
Pathological process, relevant research has been oozed and has been lured every field biology.Research shows, herbal polysaccharide have immunomodulating, antitumor,
The several functions such as antiviral, antioxidation, blood sugar lowering.Polysaccharide can promote the generation of antibody.Document report lentinan can increase
The generation of all monocyte antibodies in human body China and foreign countries, Schizophyllum commune Fr polysaccharides can promote the cell that patient's spleen forms antibody to increase.Coriolous Dersicolor (Fr.) Quel
Polysaccharide can make the patient that antibody declines produce antibody so as to immunocompetence returns to normal level.The extracting method of polysaccharide at present
Mainly there is multiple method such as decoction and alcohol sedimentation technique, membrane filter method, column chromatography, microwave loss mechanisms.
Content of the invention
Technical problem to be solved: Methods of Polysaccharide From Agrocybe Chaxingu as a kind of active component, at present with regard to the separation of Methods of Polysaccharide From Agrocybe Chaxingu
Purification process is also more, but the purity of polysaccharide that many counting methods purification obtains is relatively low, and purity disclosure satisfy that the agrocyb eaegerita of needs is many
The yield of sugar again relatively low it is impossible to the purity of Methods of Polysaccharide From Agrocybe Chaxingu can also be improved while ensureing yield it is therefore desirable to a kind of new
The extraction separation method of Methods of Polysaccharide From Agrocybe Chaxingu.
Technical scheme: for the problems referred to above, the invention discloses a kind of extraction separation method of Methods of Polysaccharide From Agrocybe Chaxingu, described
The extraction separation method step of Methods of Polysaccharide From Agrocybe Chaxingu is as follows: (1) carries out pre-cooling after fresh agrocyb eaegerita is cut into slices, and treats that agrocyb eaegerita freezes
Carry out lyophilization again after knot；(2) after lyophilization, agrocyb eaegerita piece is pulverized rapidly, added toward in agrocybe cylindracea powder after pulverizing
Ethanol water, carries out ultrasonic agitation extraction, after extraction is centrifuged extracting solution, takes supernatant after centrifugation；(3) by supernatant
Concentrated, be concentrated into 2 times that weight is agrocyb eaegerita weight；(4) agrocyb eaegerita concentrated solution is carried out membrane filtration, by molecular weight be
1000 film, collects trapped fluid；(5) film that trapped fluid is 200 by molecular weight, collects trapped fluid；(6) cutting step (5)
Liquid is stayed to be concentrated into solvent-free；(7) concentrate is carried out crystallization purifying with ethanol, crystallization time is 3h, and crystallization and filtration, after filtration
Spontaneously dry, be Methods of Polysaccharide From Agrocybe Chaxingu.
Preferably, the extraction separation method of described a kind of Methods of Polysaccharide From Agrocybe Chaxingu, the extraction of described Methods of Polysaccharide From Agrocybe Chaxingu separates
Method and step is as follows:
(1) carry out pre-cooling after fresh agrocyb eaegerita being cut into slices, carry out lyophilization after agrocyb eaegerita freezes again, freezing is dry
Dry temperature is -20 DEG C, and sublimation drying is 12h-20h；(2) after lyophilization, agrocyb eaegerita piece is pulverized rapidly, pulverized
Add the ethanol water of 20wt%-30wt% afterwards toward in agrocybe cylindracea powder, the weight of ethanol water and agrocyb eaegerita ratio is for 5:1-10:
1, carry out ultrasonic agitation extraction, ultrasonic power is 200w-400w, speed of agitator is 100rpm, extraction time is 2h-5h, extracts
Temperature is 45 DEG C, after extraction is centrifuged extracting solution, and offline rotating speed is 3500rpm, takes supernatant after centrifugation；(3) by supernatant
Liquid is concentrated, and is concentrated into 2 times that weight is agrocyb eaegerita weight；(4) agrocyb eaegerita concentrated solution is carried out membrane filtration, by molecular weight
Film for 1000, collects trapped fluid；(5) film that trapped fluid is 200 by molecular weight, collects trapped fluid；(6) by step (5)
Trapped fluid be concentrated into solvent-free；(7) concentrate is carried out crystallization purifying with ethanol, ethanol weight is concentrate weight
3-5 times of amount, crystallization temperature is 5 DEG C, and crystallization time is 3h, and crystallization and filtration spontaneously dries after filtration, is Methods of Polysaccharide From Agrocybe Chaxingu.
Preferably, in a kind of described extraction separation method of Methods of Polysaccharide From Agrocybe Chaxingu, ethanol weight is 4 times of concentrate weight.
Preferably, in a kind of described extraction separation method of Methods of Polysaccharide From Agrocybe Chaxingu, ultrasonic power is 300w.
Preferably, in a kind of described extraction separation method of Methods of Polysaccharide From Agrocybe Chaxingu ethanol water and agrocyb eaegerita weight ratio
For 7:1.
Preferably, in a kind of described extraction separation method of Methods of Polysaccharide From Agrocybe Chaxingu, ethanol water concentration is 20wt%.
Preferably, in a kind of described extraction separation method of Methods of Polysaccharide From Agrocybe Chaxingu, extraction time is 4h.
Beneficial effect: the extraction separation method of the Methods of Polysaccharide From Agrocybe Chaxingu of the present invention, the extraction process to agrocyb eaegerita and separation work
Skill is improved, and using ethanol water, Methods of Polysaccharide From Agrocybe Chaxingu is extracted, and can also carry out while ensureing extraction ratio
Remove impurity, is separated to Methods of Polysaccharide From Agrocybe Chaxingu further additionally by membrane filtration, finally by the mode of alcohol crystal purification,
Ensure while yield, to also improve the purity of the Methods of Polysaccharide From Agrocybe Chaxingu preparing.
Specific embodiment
Embodiment 1
(1) carry out pre-cooling after fresh agrocyb eaegerita being cut into slices, carry out lyophilization after agrocyb eaegerita freezes again, freezing is dry
Dry temperature is -20 DEG C, and sublimation drying is 20h；(2) after lyophilization, agrocyb eaegerita piece is pulverized rapidly, past after pulverizing
The ethanol water of 20wt% is added, the weight of ethanol water and agrocyb eaegerita ratio for 7:1, carries out ultrasonic agitation in agrocybe cylindracea powder
Extract, ultrasonic power is 300w, speed of agitator is 100rpm, extraction time is 4h, Extracting temperature is 45 DEG C, will extract after extraction
Liquid is centrifuged, and offline rotating speed is 3500rpm, takes supernatant after centrifugation；(3) supernatant is concentrated, being concentrated into weight is
2 times of agrocyb eaegerita weight；(4) agrocyb eaegerita concentrated solution is carried out membrane filtration, the film being 1000 by molecular weight, collect trapped fluid；
(5) film that trapped fluid is 200 by molecular weight, collects trapped fluid；(6) trapped fluid of step (5) is concentrated into solvent-free；(7)
Concentrate is carried out crystallization purifying with ethanol, ethanol weight is 4 times of concentrate weight, crystallization temperature is 5 DEG C, crystallization time is
3h, crystallization and filtration, spontaneously dry after filtration, be Methods of Polysaccharide From Agrocybe Chaxingu.
The Methods of Polysaccharide From Agrocybe Chaxingu purity that embodiment 1 finally gives is 95%, and final yield is 73%.
Embodiment 2
(1) carry out pre-cooling after fresh agrocyb eaegerita being cut into slices, carry out lyophilization after agrocyb eaegerita freezes again, freezing is dry
Dry temperature is -20 DEG C, and sublimation drying is 20h；(2) after lyophilization, agrocyb eaegerita piece is pulverized rapidly, past after pulverizing
The ethanol water of 20wt% is added, the weight of ethanol water and agrocyb eaegerita ratio for 10:1, carries out ultrasonic agitation in agrocybe cylindracea powder
Extract, ultrasonic power is 400w, speed of agitator is 100rpm, extraction time is 2h, Extracting temperature is 45 DEG C, will extract after extraction
Liquid is centrifuged, and offline rotating speed is 3500rpm, takes supernatant after centrifugation；(3) supernatant is concentrated, being concentrated into weight is
2 times of agrocyb eaegerita weight；(4) agrocyb eaegerita concentrated solution is carried out membrane filtration, the film being 1000 by molecular weight, collect trapped fluid；
(5) film that trapped fluid is 200 by molecular weight, collects trapped fluid；(6) trapped fluid of step (5) is concentrated into solvent-free；(7)
Concentrate is carried out crystallization purifying with ethanol, ethanol weight is 3 times of concentrate weight, crystallization temperature is 5 DEG C, crystallization time is
3h, crystallization and filtration, spontaneously dry after filtration, be Methods of Polysaccharide From Agrocybe Chaxingu.
The Methods of Polysaccharide From Agrocybe Chaxingu purity that embodiment 1 finally gives is 89%, and final yield is 77%.
Embodiment 3
(1) carry out pre-cooling after fresh agrocyb eaegerita being cut into slices, carry out lyophilization after agrocyb eaegerita freezes again, freezing is dry
Dry temperature is -20 DEG C, and sublimation drying is 12h；(2) after lyophilization, agrocyb eaegerita piece is pulverized rapidly, past after pulverizing
The ethanol water of 30wt% is added, the weight of ethanol water and agrocyb eaegerita ratio for 5:1, carries out ultrasonic agitation in agrocybe cylindracea powder
Extract, ultrasonic power is 200w, speed of agitator is 100rpm, extraction time is 5h, Extracting temperature is 45 DEG C, will extract after extraction
Liquid is centrifuged, and offline rotating speed is 3500rpm, takes supernatant after centrifugation；(3) supernatant is concentrated, being concentrated into weight is
2 times of agrocyb eaegerita weight；(4) agrocyb eaegerita concentrated solution is carried out membrane filtration, the film being 1000 by molecular weight, collect trapped fluid；
(5) film that trapped fluid is 200 by molecular weight, collects trapped fluid；(6) trapped fluid of step (5) is concentrated into solvent-free；(7)
Concentrate is carried out crystallization purifying with ethanol, ethanol weight is 5 times of concentrate weight, crystallization temperature is 5 DEG C, crystallization time is
3h, crystallization and filtration, spontaneously dry after filtration, be Methods of Polysaccharide From Agrocybe Chaxingu.
The Methods of Polysaccharide From Agrocybe Chaxingu purity that embodiment 1 finally gives is 91%, and final yield is 76%.
Claims (6)
1. a kind of extraction separation method of Methods of Polysaccharide From Agrocybe Chaxingu is it is characterised in that the extraction separation method of described Methods of Polysaccharide From Agrocybe Chaxingu
Step is as follows: (1) carries out pre-cooling after fresh agrocyb eaegerita is cut into slices, and carries out lyophilization after agrocyb eaegerita freezes again, freezing
Baking temperature is -20 DEG C, and sublimation drying is 12h-20h；(2) after lyophilization, agrocyb eaegerita piece is pulverized rapidly, powder
After broken toward agrocybe cylindracea powder in addition 20wt%-30wt% ethanol water, the weight of ethanol water and agrocyb eaegerita ratio is for 5:1-
10:1, carries out ultrasonic agitation extraction, and ultrasonic power is 200w-400w, and speed of agitator is 100rpm, and extraction time is 2h-5h, carries
Take temperature to be 45 DEG C, after extraction, extracting solution is centrifuged, centrifugal rotational speed is 3500rpm, takes supernatant after centrifugation；(3) will be upper
Clear liquid is concentrated, and is concentrated into 2 times that weight is agrocyb eaegerita weight；(4) agrocyb eaegerita concentrated solution is carried out membrane filtration, by dividing
Son measures the film for 1000, collects trapped fluid；(5) film that trapped fluid is 200 by molecular weight, collects trapped fluid；(6) by step
(5) trapped fluid is concentrated into solvent-free；(7) concentrate is carried out crystallization purifying with ethanol, ethanol weight is concentrate weight
3-5 times, crystallization temperature is 5 DEG C, and crystallization time is 3h, and crystallization and filtration spontaneously dries after filtration, is Methods of Polysaccharide From Agrocybe Chaxingu.
2. a kind of extraction separation method of the Methods of Polysaccharide From Agrocybe Chaxingu according to claim 1 is it is characterised in that described Camellia sinensis
In the extraction separation method step (7) of mushroom polysaccharide, ethanol weight is 4 times of concentrate weight.
3. a kind of extraction separation method of the Methods of Polysaccharide From Agrocybe Chaxingu according to claim 1 is it is characterised in that described Camellia sinensis
In the extraction separation method step (2) of mushroom polysaccharide, ultrasonic power is 300w.
4. a kind of extraction separation method of Methods of Polysaccharide From Agrocybe Chaxingu according to claim 1 is it is characterised in that described agrocyb eaegerita
In the extraction separation method step (2) of polysaccharide, the weight of ethanol water and agrocyb eaegerita is than for 7:1.
5. a kind of extraction separation method of the Methods of Polysaccharide From Agrocybe Chaxingu according to claim 1 is it is characterised in that described Camellia sinensis
In the extraction separation method step (2) of mushroom polysaccharide, ethanol water concentration is 20wt%.
6. a kind of extraction separation method of Methods of Polysaccharide From Agrocybe Chaxingu according to claim 1 is it is characterised in that described agrocyb eaegerita
In the extraction separation method step (2) of polysaccharide, extraction time is 4h.
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| CN108403729B (en) * | 2018-05-14 | 2021-09-14 | 广东省微生物研究所（广东省微生物分析检测中心） | Preparation method of agrocybe cylindracea extract and application of agrocybe cylindracea extract in preparation of uric acid reducing medicine |
| CN109180832A (en) * | 2018-10-12 | 2019-01-11 | 绩溪县徽菜宝生物科技有限公司 | A kind of efficient Extraction Methods of Polysaccharides From Agrocybe aegerita |
| CN110100975A (en) * | 2019-05-06 | 2019-08-09 | 四川省畜牧科学研究院 | A kind of chicken feed addictive and preparation method thereof for preventing and treating diarrhea |
| CN112079940B (en) * | 2020-09-15 | 2022-05-31 | 贵州省生物研究所 | Enrichment and purification method and application of agrocybe cylindracea polysaccharide with blood sugar reducing function |
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Inventor after: Li Zhaojia Inventor before: Wu Lei |
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Effective date of registration: 20171215 Address after: 529321 Guangdong city of Jiangmen province Kaiping city Sha Gang Industrial Park of Kaiping Ring Road No. 38 Patentee after: Kaiping Lishi Industrial Development Co., Ltd. Address before: 215000 Suzhou high tech Zone, Jiangsu International Education Park, North Road, No. 287 Patentee before: Suzhou Institute of Trade & Commerce |