CN101619107B - Astragalus polysaccharide extraction method - Google Patents

Astragalus polysaccharide extraction method Download PDF

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CN101619107B
CN101619107B CN2009101017776A CN200910101777A CN101619107B CN 101619107 B CN101619107 B CN 101619107B CN 2009101017776 A CN2009101017776 A CN 2009101017776A CN 200910101777 A CN200910101777 A CN 200910101777A CN 101619107 B CN101619107 B CN 101619107B
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astragalus
extraction
add
filter cake
ethanol
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CN101619107A (en
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陈剑英
傅锦浩
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HANGZHOU KEHUANG ENTERPRISE CO Ltd
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Abstract

The invention discloses an astragalus polysaccharide extraction method, comprising the following steps: adding ethanol to soak astragalus root powder; adding water to carry out thermal extraction; carrying out the enzymolysis of filter cakes and enzyme destruction; concentrating extract, depositing the extract with ethanol, drying the extract and the like. Compared with the traditional method, by using ethanol solution as strippant, taking water as extraction solvent, and adopting desorbing-internal boiling method, the method provided by the invention has significant improvement in extraction speed and extraction ratio, keeps the activity of the astragalus polysaccharide, and has the advantages of short time consumption, high extraction rate, less impurity and low energy consumption in the extraction process, therefore, the method is a green, safe and efficient . The method is also suitable to the extraction and the preparation of other plant-derived medicine polysaccharide.

Description

The extraction of Astragalus Polysaccharides in Astragalus method
Technical field
The present invention relates to a kind of extracting method of vegetable polysaccharides, especially a kind of method of from Chinese medicine astragalus, extracting astragalus polysaccharides.
Background technology
Along with the large-scale breeding of livestock and poultry, its stocking density improves constantly.For the generation of the disease incident, prevention and the control disease that reduce livestock and poultry, a large amount of microbiotic and the use of antiviral in aquaculture have caused the residual quantity of animal products Chinese traditional medicine to exceed standard.Have chemical sproof pathogenic micro-organism and constantly grow, caused the continuous decline of animal autoimmunity, this has caused massive losses to livestock industry.
Current, this safety of immunotherapy, green methods of treatment have become the target that worldwide insider seeks jointly.Because of many herbal medicine have the effect that improves autoimmunity, therefore, safety, Chinese herbal medicine extract efficient, low toxicity are paid close attention to by people just day by day.
Immunotherapy is a kind of notion of utilizing immunity system to treat disease.Immunotherapy utilizes immune identification, attack bacteria, exotic and paracytic function to resist and treat disease by the immunological competence of enhancing body, and wherein the polysaccharide immunotherapy has caused the great attention of various countries.
Since the seventies in 20th century, scientists finds that constantly polysaccharide and saccharide complex not only exist as the material of the energy and formation organism in organism, and it also is present in all membrane structures, participates in the life movement of various types of cells, has diversified biological function.
Polysaccharide is the quite complicated macromolecular compound of a kind of structure.Biological activity and the functions complexity that it had, multipath, many environment, many target spots are called as the biological response effector at international academic community.Based on dextran, be to prop up special space structure of chain formation in the composition of polysaccharide, make it have good physiologically active by β-1,3 glucoside bond bonded straight chain and β-1,6 glucoside bond bonded.
The Radix Astragali that is called " ten medicines, eight stilbenes " by pharmaceutical industry has the effect of tonifying Qi and lifting yang, strengthening superficial resistance to stop perspiration, inducing diuresis to remove edema, apocenosis, expelling pus and promoting granulation, has effect anti-oxidant, antiviral, that alleviate immunologic injury and immune two-ways regulation.Tradition is used to improve effects such as body immunity.Astragalus polysaccharides is one of the most effective composition in the Radix Astragali, has good water-solubility.
Existing extraction of Astragalus Polysaccharides in Astragalus method mainly contains water and carries and sending out and microwave method.The water extraction extraction yield is low, and the step complexity, length consuming time, and loss of effective components is more; Microwave method is higher to equipment requirements, implements difficulty, is difficult to large-scale application.
Summary of the invention
The purpose of this patent provides a kind of extraction of Astragalus Polysaccharides in Astragalus method, is main raw material with plant-derived Chinese medicine astragalus, handles through two step of desorb-inner ebullition extraction process, obtains high-level efficiency, high-load astragalus polysaccharides, can be used for strengthening fowl poultry autoimmunity.
Above-mentioned purpose of the present invention is achieved through the following technical solutions:
A kind of extraction of Astragalus Polysaccharides in Astragalus method is characterized in that may further comprise the steps:
1) in the astragalus root powder, adds ethanol, leave standstill 30min~60min;
2) with the amount of 3~5 times of astragalus root grain weight amounts, the adding temperature is 60~100 ℃ a water, insulation, backflow 15min~30min, and cooling is filtered, and water cleans repeatedly, and filtrate and scavenging solution merge, and filter cake is standby;
3) add the water of 1~10 times of weight toward above-mentioned filter cake, add zymin, controlled temperature is 40~60 ℃, and it be neutral regulating pH, and enzymolysis 1~8h post-heating enzyme that goes out that heats up is cooled to room temperature, filter, and cleaning, filter cake discards, merging filtrate and scavenging solution;
4) merge 2,3 step gained filtrate and scavenging solutions, be evaporated to 10%~50% of original volume, add 95% ethanol and carry out alcohol precipitation, cross filter solid, drying gets the astragalus polysaccharides product.
Further, described enzyme is one or more in neutral cellulase, neutral protease or the papain, is preferably neutral cellulase.The add-on of enzyme is 2 ‰~5% of an astragalus root powder.
Further, the alcoholic acid add-on is 1.5~3 times of astragalus root grain weight amount in the step 1; Soaking time in the step 2 is preferably 25min; Enzymolysis time in the step 3 is preferably 2h.
The present invention is directed to this botanical herbs material of the Radix Astragali, adopt the extracting method of desorb-inner ebullition method (abbreviation two-step approach).
It is strippant that the inventive method is selected ethanolic soln for use, water is for extracting solvent, utilize a small amount of strippant of being added in the first step desorption process lower boiling vaporization medium cleverly as material Milkvetch Root inside, make its second step add fast heat extract solvent after, instant vaporization under the situation that has little time to spread in material inside, thereby make the inner mass transfer mode of material that change take place.Effective constituent changes the convection current diffusion into by original common molecular diffusion, thereby makes extraction rate obviously accelerate, and the easier stripping of polysaccharide.
Enzymolysis process can extract on the one hand the extraction of the polysaccharide in the fiber of residual existence in the filter cake, can also remove impurity such as protein, amino acid on the other hand, improves the purity of astragalus polysaccharides.
The present invention adopts two-step approach, compared to traditional water extraction and microwave method, its extraction rate and extraction yield all are significantly improved, and the activity that has kept astragalus polysaccharides, whole leaching process weak point consuming time, the yield height, impurity is few, power consumption is low, be a kind of green, safety, extraction of Astragalus Polysaccharides in Astragalus method efficiently.The inventive method also is applicable to the extraction and the preparation of other plant source property medicine polysaccharide.
Embodiment
Below in conjunction with specific embodiment content of the present invention is further described.
Embodiment 1
Take by weighing 70g astragalus root powder and place the 500ml there-necked flask, dripping 140ml concentration is 70% ethanol, makes material evenly wetting, leaves standstill 30min.
Add the 245ml temperature in the above-mentioned material and be 100 ℃ distilled water,, cooling, filter with constant temperature sleeve insulation, backflow 20min; Filter cake washes repeatedly with 40 ℃ of hot water, mixes filtrate and scavenging solution, and filter cake is standby.
Water toward 5 times of weight of above-mentioned filter cake adding adds the 0.35g neutral cellulase, and controlled temperature is 48 ℃, and it is neutral regulating pH, enzymolysis 2h.Enzymolysis finishes the post-heating material and is warming up to 80 ℃, the enzyme that goes out, and cooling, filtration, cleaning, merging filtrate and scavenging solution, filter cake discards.
The filtrate and the scavenging solution that merge above step gained in the step, be evaporated to 20% of original volume, alcohol precipitation is carried out with 95% ethanol in the cooling back, the gained throw out filters, with protein residual in the acetone washing and precipitating, Mierocrystalline cellulose and part moisture content, final drying obtains astragalus polysaccharides 11.15g, and calculating its yield is 15.9%.
Embodiment 2
Take by weighing 50g astragalus root powder and place the 500ml there-necked flask, dripping 75ml concentration is 95% ethanol, makes material evenly wetting, leaves standstill 30min.
Add the 250ml temperature in the above-mentioned material and be 100 ℃ distilled water,, cooling, filter with constant temperature sleeve insulation, backflow 20min; Filter cake washes repeatedly with 40 ℃ of hot water, mixes filtrate and scavenging solution, and filter cake is standby.
Water toward 10 times of weight of above-mentioned filter cake adding adds the 0.5g neutral cellulase, and controlled temperature is 50 ℃, and it is neutral regulating pH, enzymolysis 5h.Enzymolysis finishes the post-heating material and is warming up to 80 ℃, the enzyme that goes out, and cooling, filtration, cleaning, merging filtrate and scavenging solution, filter cake discards.The filtrate and the scavenging solution that merge above step gained in the step, be evaporated to 20% of original volume, alcohol precipitation is carried out with 95% ethanol in the cooling back, the gained throw out filters, with protein residual in the acetone washing and precipitating, Mierocrystalline cellulose and part moisture content, final drying obtains astragalus polysaccharides 8.84g, and calculating its yield is 17.7%.
Embodiment 3
Take by weighing 65g astragalus root powder and place the 500ml there-necked flask, dripping 140ml concentration is 70% ethanol, makes material evenly wetting, leaves standstill 30min.
Add the 200ml temperature in the above-mentioned material and be 90 ℃ distilled water,, cooling, filter with constant temperature sleeve insulation, backflow 20min; Filter cake washes repeatedly with 40 ℃ of hot water, mixes filtrate and scavenging solution, and filter cake is standby.
Water toward 5 times of weight of above-mentioned filter cake adding adds the 0.46g neutral cellulase, and controlled temperature is 48 ℃, and it is neutral regulating pH, enzymolysis 2h.Enzymolysis finishes the post-heating material and is warming up to 80 ℃, the enzyme that goes out, and cooling, filtration, cleaning, merging filtrate and scavenging solution, filter cake discards.The filtrate and the scavenging solution that merge above step gained in the step, be evaporated to 20% of original volume, alcohol precipitation is carried out with 95% ethanol in the cooling back, the gained throw out filters, with protein residual in the acetone washing and precipitating, Mierocrystalline cellulose and part moisture content, final drying obtains astragalus polysaccharides 9.34g, and calculating its yield is 14.4%.
Embodiment 4
Take by weighing 65g astragalus root powder and place the 500ml there-necked flask, dripping 140ml concentration is 70% ethanol, makes material evenly wetting, leaves standstill 30min.
Add the 280ml temperature in the above-mentioned material and be 70 ℃ distilled water,, cooling, filter with constant temperature sleeve insulation, backflow 20min; Filter cake washes repeatedly with 40 ℃ of hot water, mixes filtrate and scavenging solution, and filter cake is standby.
Water toward 5 times of weight of above-mentioned filter cake adding adds the 0.33g neutral cellulase, and controlled temperature is 48 ℃, and it is neutral regulating pH, enzymolysis 2h.Enzymolysis finishes the post-heating material and is warming up to 80 ℃, the enzyme that goes out, and cooling, filtration, cleaning, merging filtrate and scavenging solution, filter cake discards.The filtrate and the scavenging solution that merge above step gained in the step, be evaporated to 20% of original volume, alcohol precipitation is carried out with 95% ethanol in the cooling back, the gained throw out filters, with protein residual in the acetone washing and precipitating, Mierocrystalline cellulose and part moisture content, final drying obtains astragalus polysaccharides 7.66g, and calculating its yield is 11.8%.
Embodiment 5
Take by weighing 60g astragalus root powder and place the 500ml there-necked flask, dripping 140ml concentration is 70% ethanol, makes material evenly wetting, leaves standstill 40min.
Add the 200ml temperature in the above-mentioned material and be 100 ℃ distilled water,, cooling, filter with constant temperature sleeve insulation, backflow 30min; Filter cake washes repeatedly with 40 ℃ of hot water, mixes filtrate and scavenging solution, and filter cake is standby.
Water toward 3 times of weight of above-mentioned filter cake adding adds the 1.8g neutral protease, and controlled temperature is 55 ℃, and it is neutral regulating pH, enzymolysis 2h.Enzymolysis finishes the post-heating material and is warming up to 80 ℃, the enzyme that goes out, and cooling, filtration, cleaning, merging filtrate and scavenging solution, filter cake discards.The filtrate and the scavenging solution that merge above step gained in the step, be evaporated to 30% of original volume, alcohol precipitation is carried out with 95% ethanol in the cooling back, the gained throw out filters, with protein residual in the acetone washing and precipitating, Mierocrystalline cellulose and part moisture content, final drying obtains astragalus polysaccharides 9.69gg, and calculating its yield is 16.2%.
Embodiment 6
Take by weighing 50g astragalus root powder and place the 500ml there-necked flask, dripping 100ml concentration is 95% ethanol, makes material evenly wetting, leaves standstill 30min.
Add the 250ml temperature in the above-mentioned material and be 100 ℃ distilled water,, cooling, filter with constant temperature sleeve insulation, backflow 25min; Filter cake washes repeatedly with 40 ℃ of hot water, mixes filtrate and scavenging solution, and filter cake is standby.
Water toward 7 times of weight of above-mentioned filter cake adding adds the 1g papain, and controlled temperature is 60 ℃, and it is neutral regulating pH, enzymolysis 3h.Enzymolysis finishes the post-heating material and is warming up to 80 ℃, the enzyme that goes out, and cooling, filtration, cleaning, merging filtrate and scavenging solution, filter cake discards.
The filtrate and the scavenging solution that merge above step gained in the step, be evaporated to 20% of original volume, alcohol precipitation is carried out with 95% ethanol in the cooling back, the gained throw out filters, with protein residual in the acetone washing and precipitating, Mierocrystalline cellulose and part moisture content, final drying obtains astragalus polysaccharides 8.42g, and calculating its yield is 16.9%.

Claims (5)

1. extraction of Astragalus Polysaccharides in Astragalus method is characterized in that may further comprise the steps:
1) add ethanol in the astragalus root powder, leave standstill 30min~60min, the alcoholic acid add-on is 1.5~3 times of astragalus root grain weight amount;
2) with the amount of 3~5 times of astragalus root grain weight amounts, the adding temperature is 60~100 ℃ a water, insulation, backflow 15min~30min, and cooling is filtered, and water cleans repeatedly, and filtrate and scavenging solution merge, and filter cake is standby;
3) add the water of 1~10 times of weight toward above-mentioned filter cake, add zymin, controlled temperature is 40~60 ℃, and it be neutral regulating pH, and enzymolysis 1~8h post-heating enzyme that goes out that heats up is cooled to room temperature, filter, and cleaning, filter cake discards, merging filtrate and scavenging solution;
4) merge 2,3 step gained filtrate and scavenging solutions, be evaporated to 10%~50% of original volume, add 95% ethanol and carry out alcohol precipitation, cross filter solid, drying gets the astragalus polysaccharides product.
2. extraction of Astragalus Polysaccharides in Astragalus method as claimed in claim 1 is characterized in that described enzyme is one or more in neutral cellulase, neutral protease or the papain, and the add-on of enzyme is 2 ‰~5% of an astragalus root powder.
3. extraction of Astragalus Polysaccharides in Astragalus method as claimed in claim 2 is characterized in that described enzyme is a neutral cellulase.
4. extraction of Astragalus Polysaccharides in Astragalus method as claimed in claim 3 is characterized in that the soaking time in the step 2 is 25min.
5. extraction of Astragalus Polysaccharides in Astragalus method as claimed in claim 4 is characterized in that the enzymolysis time in the step 3 is 2h.
CN2009101017776A 2009-08-17 2009-08-17 Astragalus polysaccharide extraction method Expired - Fee Related CN101619107B (en)

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CN102850463A (en) * 2012-09-28 2013-01-02 江西田昌农业技术开发有限公司 Preparation method of allium chinense polysaccharide
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CN103316070A (en) * 2013-07-10 2013-09-25 吴中区胥口精益生物医药研究所 Radix astragali essence as well as preparation method and pharmaceutical application thereof
CN103382228A (en) * 2013-07-31 2013-11-06 瑞普(天津)生物药业有限公司 Preparation method for high-activity astragalus polysaccharide
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CN104719632A (en) * 2013-12-19 2015-06-24 天津工业大学 Feed additive and preparation method thereof
CN103980375A (en) * 2014-05-20 2014-08-13 河南中烟工业有限责任公司 Astragalus polysaccharide, extracting and purifying method and application thereof as tobacco moistening agent
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