CN109503729A - A method of extracting polysaccharide - Google Patents
A method of extracting polysaccharide Download PDFInfo
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- CN109503729A CN109503729A CN201811514029.6A CN201811514029A CN109503729A CN 109503729 A CN109503729 A CN 109503729A CN 201811514029 A CN201811514029 A CN 201811514029A CN 109503729 A CN109503729 A CN 109503729A
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- polysaccharide
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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
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Abstract
The invention discloses a kind of method for extracting polysaccharide, include the following steps: that, so that polysaccharide is dissolved in the water, cooling, filtering obtains filtered fluid Step 1: the substance containing polysaccharide is impregnated in water；Step 2: freezing and filtering liquid to filtered fluid freezes；Step 3: the filtered fluid of icing is thawed, filtering obtains filter residue, and dry filter residue obtains polysaccharide.Step of the present invention is simple, and Polyose extraction is convenient, does not need using organic solvent, therefore will not generate pollution, and the polysaccharide extracted is pure, does not need accurate solution proportion, is suitable for large-scale industrial production.
Description
Technical field
The invention belongs to drug field, in particular to a kind of method for extracting polysaccharide.
Background technique
Polysaccharide (polysaccharide) is the sugar chain combined by glycosidic bond, to be at least more than that 10 monosaccharide forms
Polymerization sugar polymeric carbohydrate.Polysaccharide it is widely used, such as:
Immunological regulation:
After Hosono Akira etc. extracts the cell Ultrasonic Pulverization of Bifidobacterium bacterium, with ultrafiltration apparatus and anion
Exchanger resin, gel chromatography are purified into the polysaccharide with immune-enhancing activity.Oka Shuichi etc. is from purple perilla (Perilla)
Isolated polysaccharide has anti-allergy action.Fujimiy hjaki is from the fructification of Agaricus (Agr/cus) plant
The polysaccharide extracted have immunosuppressive action, it can be reduced our usually used immunosuppressor such as cytotoxicity,
Body anti-infection ability declines, to side effects such as the Reproduction suppressions of myeloid element, this polysaccharide can be made oral or injection
With drug, a kind of functional food may be made as.
Antiviral and anticancer:
The Antiviral Mechanism of most of polysaccharide is the absorption for inhibiting virus to cell, this may be due to polysaccharide macro-molecular machine
Tool is chemically integrated on the Gp120 molecule of HW-I, the binding site of virus with cell has been covered, thus competitively
Virus infected cell is blocked.Hara Masahiko etc. obtains a kind of phytopathy from the tealeaves that the first season and the second season harvest
Malicious inhibitor, it is a kind of monosaccharide or polysaccharide component containing tannic acid, not only can inhibit the pathogenic effects of virus, but also can be pressed down
Make the propagation of virus.It is reported that also isolated can largely have anticancer from the culture of Agaricus (Agr/cus) plant
The water-soluble substances of active constituent, including the acidic polysaccharose, water-soluble neutral more isolated from the fructification of mushroom platymiscium
Sugar and water-solubility protein polysaccharide.NodaKiyoshiC and Kato Toshimitsu etc. is separated from chlorella and spirulina respectively
The polysaccharide and controlling sulfate polyose for providing anticancer activity, can inhibit metastases, safety better than traditional operative treatment and
Chemotherapy.Nakano Masa hi is from the nut of pecan, Cortex Eucommiae and leguminous plant Aspa/athus linearis
A kind of anti-oxidant acidic polysaccharose is extracted, it can not only inhibit the duplication of the retrovirus such as AIDS virus, and can rise
To immunoregulation effect, alternative traditional biggish antiviral drugs of expensive and side effect to a certain extent.
It is hypoglycemic:
Ukai Shigeo etc. extracts the acidity of antihyperglycemic from a kind of fructification of tremella (KINJI) or mycelium
Polysaccharide.FujiiMakoto etc. extracts a kind of algal polysaccharides that can reduce blood glucose level from seaweed plant, and is made
With fucose (Fucoidan) health food as main component, it can significantly improve the immune function of people.
Treatment:
The Radix Salviae Miltiorrhizae polysaccharide that Kanou Kokuki etc. is isolated from Radix Salviae Miltiorrhizae is able to suppress the secretion of Urine proteins, alleviates liver kidney disease
Disease symptoms can be made into oral or intramuscular formulation, reduces and makes due to taking the steroids such as Dipyridamole or platelet suppressant drug for a long time
At adverse reaction.ShibatHideyuki etc. has invented a kind of polysaccharide system containing sulphation fucose active constituent
Agent, it can be reduced the side effect of the Nonsteroidal anti-inflammatories analgestic such as indocin, aspirin.
Beauty:
Honda Yasuki etc. obtains one kind from Common Primrose category (Polyanthus) plant to be had good moisturizing, resists
The acid heteroglycan of the effects of wrinkle.Sawai Yasuko etc. is isolated from the rhizome of rhizoma acori graminei (Acorusgram/neus)
Polysaccharide can inhibit the generation of melanin, there is anti-inflammatory, antioxidation, can be used for the treatment of melanose, and because it is with good
Good moisture-keeping function, therefore but also as the effective component of cosmetics.The meat from shellfish such as Shimomura K~nji
A kind of acidic polysaccharose with beauty functions is obtained in catabolite.It is demonstrated experimentally that this acidic polysaccharose can inhibit anti-aging
The decomposition of hyaluronic acid reduces skin lines and dry and cracked, thus can be used as the effective component of beauty food and cosmetics.
Emulsifier:
Keiichi etc. is mentioned from the body cell wall of grass family (Gramineae) Festuca (Festuca) plant (such as barley)
The polysaccharide with emulsification is obtained, emulsifier is can be used as and is widely used in industrial production, and is safe and pollution-free.
KuraneRyuichiro etc. is obtained by culture Alcaligenes latus B-16 (~3ca//geneshuus B-16) and is isolated one
The polysaccharide that kind is made of trehalose and mannose, dissolubility is good in water for this polysaccharide, has good stability, can be used as grinding
Agent, emulsifier stabilizer and thickener.
Other purposes:
Sakata Shigenobu etc. is by the way that a variety of monosaccharide, polysaccharide and its derivatization carbohydrate, (such as aldose sticks polysaccharide, more
Sugar after glycolysis) it is fermented or is extracted, the reagent of first stability, safety is obtained, it can reduce typical nuisance (such as two
Oxygen glutinous rehmannia, cyano compound, Polychlorinated biphenyls etc.) infringement to environment and human body is very meaningful environmentally friendly reagent.Watanabe
Sa J one kind is to adsorb the culture substrate culture osteoblast that the hydroxyapatite of polysaccharide (such as starch) makees carrier.The spy of this carrier
Point is that osteoblast growth factor receptors can be stimulated by not having to be added the substances such as serum, Porcine HGF, and it can keep away
Exempt from when cultivating certain osteoblast, as serum type specificity and a large amount of people spent by most suitable serum must be screened
Power, financial resources, thus the appearance of the present invention undoubtedly greatly reduces the culture expense of osteoblast, has high economic valence
Value and social value.
But the existing method that polysaccharide is prepared from plant is using alcohol deposition method, i.e., in the aqueous solution for being dissolved with polysaccharide
Middle addition ethyl alcohol comes out polysaccharide precipitation to reduce the solubility of polysaccharide.The shortcomings that this method are as follows: one, in addition to polysaccharide
Outside, the substances such as DNA, protein can also be precipitated as concentration of alcohol reduces, and cause the polysaccharide impurity extracted excessive；Two, it needs
Using ethyl alcohol, and need the accurate proportion of second alcohol and water, it is also desirable to mix, therefore complicated operation, be not suitable for high-volume
The industrial abstract of pipelining；Three, contain ethyl alcohol in waste liquid, it is not easy to handle, arbitrarily topple over and pollutes the environment.
Summary of the invention
In order to overcome the above problems, the present invention provides a kind of methods for extracting polysaccharide.Step of the present invention is simple, and polysaccharide mentions
It takes conveniently, does not need using organic solvent, therefore pollution will not be generated, and the polysaccharide extracted is pure, does not need accurate solution and matches
Than being suitable for large-scale industrial production.
The technical solution adopted by the present invention to solve the technical problems is:
A method of polysaccharide is extracted, is included the following steps:
Step 1: the substance containing polysaccharide is impregnated in water, so that polysaccharide is dissolved in the water, cooling, filtering is obtained
Filtrate；
Step 2: freezing and filtering liquid to filtered fluid freezes；
Step 3: the filtered fluid of icing is thawed, filtering obtains filter residue, and dry filter residue obtains polysaccharide.
Further to improve, so that the method that polysaccharide is dissolved in the water is reflux extraction, refluxing extraction is to when setting
Between after, cooling, filtering obtains filtered fluid
Further to improve, the temperature of refluxing extraction is 40-100 DEG C, and the time is -3 hours 0.5 hour；The water is to steam
Distilled water or containing acid or aqueous solution of alkali etc..
Further to improve, in the step 1, the substance containing polysaccharide is Chinese medicine.
Further to improve, the Chinese medicine includes Ma Kalie third, Sheepear Inula Herb, Poria cocos, fructus lycii, dendrobium nobile, ganoderma lucidum and folium artemisiae argyi
Deng.
Further to improve, the mass ratio of the substance and water containing polysaccharide is 1-1:1-100.
Further to improve, the mass ratio of the substance and water containing polysaccharide is 1:50.
Further to improve, in the step 3, the time that the filtered fluid of icing thaws is 0.5-24 hours；It thaws
Temperature is 0-40 DEG C.
Further to improve, the method that polysaccharide is dissolved in the water is ultrasonic extraction, the frequency of ultrasonic wave
For 25KHZ-130KHZ, time 0.1-3h.
Detailed description of the invention
Fig. 1 is molish response diagram；
Fig. 2 is UV absorption figure of the precipitating after phenol sulfuric acid develops the color；
Fig. 3 is the standard curve of glucose；
Fig. 4 compares figure for the purity of new method and alcohol extracting method.
Specific embodiment
The present invention is further elaborated and is illustrated with Figure of description combined with specific embodiments below:
Embodiment one
1. the preparation and identification of polysaccharide
The third 3 parts of Ma Kalie are taken, every part of 1g is accurately weighed, and the distilled water of 50ml is added, and refluxing extraction 1h is stood, and room temperature is cold
But, it filters, takes subsequent filtrate, be respectively placed in -4 DEG C, -20 DEG C, -80 DEG C of freezings freeze, and take out afterwards for 24 hours, thaw at RT, there have to be heavy
Precipitation goes out.
Molish reaction can occur for precipitating, and such as Fig. 1 illustrates that this is precipitated as polysaccharide.
For precipitating after phenol sulfuric acid develops the color, ultraviolet absorption curve illustrates this as shown in Fig. 2, a length of 490nm of maximum absorption wave
It is precipitated as polysaccharide.
2. the recovery rate and purity of polysaccharide difference preparation method
2.1. the recovery rate of polysaccharide
The third 4 parts of Ma Kalie are taken, every part of 5g is accurately weighed, and the distilled water of 250ml is added, and refluxing extraction 1h is stood, room temperature
Cooling, filtering takes subsequent filtrate, takes wherein 2 parts, is placed in -80 DEG C of freezings and freezes, takes out afterwards for 24 hours, after thaw at RT, filters, does
Dry, the quality of accurately weighed precipitating calculates recovery rate.
Other 2 parts, it is slowly added to 95% ethyl alcohol, makes final determining alcohol 80%, after standing 8h, is filtered, dry, precision claims
Surely the quality precipitated calculates recovery rate.
New method, polysaccharide average recoveries %=12.08% after freeze-thaw
Conventional method, polysaccharide average recoveries %=17.62% after water extract-alcohol precipitation
Illustrate: water extraction and alcohol precipitation method recovery rate is higher than new method.
2.2. the purity of polysaccharide
2.2.1. the foundation of standard curve
Accurately weighed glucose control product 1mg, as in the volumetric flask of 100ml, with distilled water constant volume to get glucose mark
Quasi- product solution.It is accurate respectively to measure 0.2,0.4,0.6,0.8,1,1.2,1.4ml reference substance solution, it is separately added into and all steams in right amount
Distilled water polishing is to 2ml, and rear that 1ml phenol is added, 5ml sulfuric acid places 10min after concussion, boiling water bath 20min is cooled to room temperature
Afterwards, with ultraviolet specrophotometer under 490nm wavelength, absorbance value is surveyed.Standard curve is as shown in Figure 3.
2.2.2. the measurement of purity
Observed under daylight is set in above two method (new method and water extraction and alcohol precipitation method) preparation, as shown in figure 4, result is preliminary
Show that the total starches of new method preparation are whiter than polysaccharide prepared by water extract-alcohol precipitation.
It is upper in the total starches of Fig. 4: the total starches of new method preparation, under: the total starches of water extract-alcohol precipitation preparation
Each 2 parts of total starches sample of above two method preparation are taken, totally 4 parts, every part of 10mg is accurately weighed, as 50ml
In volumetric flask, with distilled water constant volume to get test solution.
Test solution 0.5ml is taken, 1.5ml distilled water, 1ml phenol is added, 5ml sulfuric acid places 10min after concussion, boil
Water-bath 20min after being cooled to room temperature, with ultraviolet specrophotometer under 490nm wavelength, surveys absorbance value.
It is 12.08%*65.92%=7.96% that i.e. new method, which finally obtains the yield of polysaccharide,；
Water extract-alcohol precipitation finally obtains the yield of polysaccharide are as follows: 17.62%*41.19%=7.26%.
Specific embodiment two
1. the recovery rate of polysaccharide
4 parts of Sheepear Inula Herb are taken, every part of 5g is accurately weighed, and the distilled water of 250ml, ultrasonic extraction 1h is added, and supersonic frequency is
25KHZ, power 500W, ultrasonic temperature are room temperature, about 10 DEG C, are stood, and filtering takes subsequent filtrate, takes wherein 2 parts, be concentrated into
25ml is placed in -80 DEG C of freezings and freezes, takes out afterwards for 24 hours, after thaw at RT, filters, dry, and the quality of accurately weighed precipitating calculates
Recovery rate.
Other 2 parts, it is slowly added to 95% ethyl alcohol, makes final determining alcohol 80%, after standing 8h, is filtered, dry, precision claims
Surely the quality precipitated calculates recovery rate.
New method, polysaccharide average recoveries %=0.81% after freeze-thaw
Conventional method, polysaccharide average recoveries %=1.98% after water extract-alcohol precipitation
Illustrate: water extraction and alcohol precipitation method recovery rate is higher than new method.
2. the measurement of the purity of polysaccharide
Each 2 parts of total starches sample of above two method preparation are taken, totally 4 parts, every part of 10mg is accurately weighed, as 50ml
In volumetric flask, with distilled water constant volume to get test solution.
Test solution 1ml is taken, 1ml distilled water, 1ml phenol is added, 5ml sulfuric acid places 10min, boiling water bath after concussion
20min after being cooled to room temperature, with ultraviolet specrophotometer under 490nm wavelength, surveys absorbance value.
It is 0.81%*45.14%=0.35% that i.e. new method, which finally obtains the yield of polysaccharide,；
Water extract-alcohol precipitation finally obtains the yield of polysaccharide are as follows: 1.98%*16.64%=0.33%.
Therefore present invention side's new method extracts the purity of polysaccharide and final yield is above traditional water extraction and alcohol precipitation method, and grasps
Make simply, to be less prone to pollution.
Test finds that extracting method of the invention needs polysaccharide to reach a certain concentration and freeze and reaches certain time, specifically
When polysaccharide concentration is the concentration of 0.731mg/ml, freezing freezes 3 hours, and rear fusing freezed 8 as a child without Precipitation
There is Precipitation；And polysaccharide concentration has Precipitation in freezing 3 hours when reaching 10mg/ml；So being needed when polysaccharide concentration is low
Suitably to increase cooling time.
Finally it should be noted that the above embodiments are merely illustrative of the technical solutions of the present invention, rather than the present invention is protected
The limitation of range is protected, although explaining in detail referring to preferred embodiment to the present invention, those skilled in the art are answered
Work as understanding, it can be with modification or equivalent replacement of the technical solution of the present invention are made, without departing from the reality of technical solution of the present invention
Matter and range.
Claims (9)
1. a kind of method for extracting polysaccharide, which comprises the steps of:
Step 1: the substance containing polysaccharide is impregnated in water, so that polysaccharide is dissolved in the water, cooling, filtering is filtered
Liquid；
Step 2: freezing and filtering liquid to filtered fluid freezes；
Step 3: the filtered fluid of icing is thawed, filtering obtains filter residue, and dry filter residue obtains polysaccharide.
2. a kind of method for extracting polysaccharide as described in claim 1, which is characterized in that so that the method that polysaccharide is dissolved in the water
For reflux extraction, refluxing extraction is to after setting time, and cooling, filtering obtains filtered fluid.
3. a kind of method for extracting polysaccharide as claimed in claim 2, which is characterized in that the temperature of refluxing extraction is 40-100
DEG C, the time is -3 hours 0.5 hour.
4. a kind of method for extracting polysaccharide as described in claim 1, which is characterized in that in the step 1, contain polysaccharide
Substance is Chinese medicine.
5. a kind of method for extracting polysaccharide as claimed in claim 4, which is characterized in that the Chinese medicine include Ma Kalie third,
Sheepear Inula Herb, Poria cocos, fructus lycii, dendrobium nobile, ganoderma lucidum and folium artemisiae argyi.
6. a kind of method for extracting polysaccharide as described in claim 1, which is characterized in that the matter of the substance and water containing polysaccharide
Amount is than being 1-1:1-100.
7. a kind of method for extracting polysaccharide as claimed in claim 6, which is characterized in that the matter of the substance and water containing polysaccharide
Amount is than being 1:50.
8. a kind of method for extracting polysaccharide as described in claim 1, which is characterized in that in the step 3, the filtering of icing
The time that lyolysis is frozen is 0.5-24 hours；The temperature of defrosting is 0-40 DEG C.
9. a kind of method for extracting polysaccharide as described in claim 1, which is characterized in that described that polysaccharide is made to be dissolved in the water
Method is ultrasonic extraction, and the frequency of ultrasonic wave is 25KHZ-130KHZ, time 0.1h-3h.
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Cited By (3)
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| CN110526999A (en) * | 2019-08-27 | 2019-12-03 | 湖南医药学院 | A kind of extracting method and application thereof of the third Thick many candies of Ma Kalie |
| CN110522781A (en) * | 2019-08-27 | 2019-12-03 | 湖南医药学院 | A kind of the third active material of Ma Kalie extracting method and application thereof |
| CN111418823A (en) * | 2020-04-27 | 2020-07-17 | 福建省泉州喜多多食品有限公司 | Glutinous-cotton white fungus soup and making method thereof |
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| CN110522781A (en) * | 2019-08-27 | 2019-12-03 | 湖南医药学院 | A kind of the third active material of Ma Kalie extracting method and application thereof |
| CN110522781B (en) * | 2019-08-27 | 2021-07-30 | 湖南医药学院 | Extraction method and application of macaroni active substance |
| CN111418823A (en) * | 2020-04-27 | 2020-07-17 | 福建省泉州喜多多食品有限公司 | Glutinous-cotton white fungus soup and making method thereof |
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