CN104031155A - Industrial extraction and purification method of Quanzhenyiqi decoction polysaccharides - Google Patents
Industrial extraction and purification method of Quanzhenyiqi decoction polysaccharides Download PDFInfo
- Publication number
- CN104031155A CN104031155A CN201410188735.1A CN201410188735A CN104031155A CN 104031155 A CN104031155 A CN 104031155A CN 201410188735 A CN201410188735 A CN 201410188735A CN 104031155 A CN104031155 A CN 104031155A
- Authority
- CN
- China
- Prior art keywords
- polysaccharide
- solution
- extraction
- purification
- polysaccharides
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Abstract
The invention discloses an industrial extraction and purification method of Quanzhenyiqi decoction polysaccharides, belongs to a Chinese traditional medicinal extraction technology, and aims to provide a simple extraction and purification method of high-content polysaccharides. The method comprises the following steps: weighing all components, and drying and crushing all the medicinal materials except prepared rehmannia root; segmenting the prepared rehmannia root, extracted through an ultrasonic extraction processing, centrifuging, and taking the obtained supernatant; concentrating the supernatant, adding ethanol until the ethanol content of the obtained medicinal liquor is 75-85%, allowing the medicinal liquor to stand, centrifuging, taking the obtained lower precipitate, and drying to obtain crude polysaccharides A; taking the crude polysaccharides A, adding water to dissolve the crude polysaccharides A, uniformly shaking, adding an acid to adjust the pH value of the obtained crude polysaccharide A solution to 5-6, adding papain, carrying out enzymatic hydrolysis, carrying out inactivation centrifugation, removing the obtained precipitate, and taking the obtained lower liquid to obtain a crude polysaccharide enzymatic hydrolysate A; adding a chloroform-n-butanol solution to the crude polysaccharide enzymatic hydrolysate A, shaking, allowing the obtained solution to stand overnight, and collecting the obtained upper water solution to obtain a polysaccharide solution A; and adding water to prepare a solution, adjusting the pH value of the obtained polysaccharide solution to 4.5-5.5, adding the polysaccharide solution to macro-porous resin for adsorption, filtering to obtain crude polysaccharides, and ultrafiltering the solution to obtain refined polysaccharides.
Description
Technical field
The present invention relates to a kind of extracting and purifying method of polysaccharide, relate in particular to a kind of method of industrial extraction and purification of a green authentic gas soup polysaccharide; Belong to traditional Chinese medicine extraction technical field.
Background technology
An authentic gas soup is that bright clear medical science Feng Zhao opens the famous prescription in " the secret record of Feng Shi embroidered purse ".Formed by Radix Rehmanniae Preparata, the bighead atractylodes rhizome, ginseng, the tuber of dwarf lilyturf, shizandra berry, monkshood, the root of bidentate achyranthes.The clinical treatment deficient property disease in various old age that is widely used in, as gastrointestinal illness, chronic obstructive pulmonary disease, lung cancer, etc. all empty immunologic hypofunction persons of dialectical genus negative and positive of qi and blood, evident in efficacy.Modern study shows, polysaccharide has the immunity of adjusting, and reducing blood-fat is antitumor, anti-inflammatory, the anti-ageing physiological function of waiting for a long time.
At present about the extraction research report of an authentic gas soup polysaccharide is little, this seminar has carried out discussion to a certain degree to an authentic gas soup polysaccharide in recent years, find that its main pharmacological action is strengthening immunity, the non-specific immune function of normal mouse and And Utilization of CTX-Immunosuppressed-mice is had to enhancement in various degree, can improve the humoral immune function that endoxan suppresses, there is Intestinal Mucosal Immunization regulating effect, its dominant mechanism is rising immunosuppressed mice blood serum IL-6 and TNF-alpha levels, immune stimulatory suppresses the cytokine secretion of mouse body, thereby reach the effect that regulates immunologic function.
Although Chen Hong provides " extraction of an authentic gas soup total polysaccharides and quality control preliminary study " in Master's thesis, the polysaccharide content of gained is not high, and extraction yield is also lower.
Summary of the invention
For above-mentioned deficiency, goal of the invention is to provide a kind of extracting and purifying method simple, the extracting and purifying method of the authentic gas soup polysaccharide that polysaccharide content is high.
For this reason, technical scheme provided by the invention is such: the method for the industrial extraction and purification of this authentic gas soup polysaccharide, comprises the steps: successively
1) take Radix Rehmanniae Preparata, the bighead atractylodes rhizome, ginseng, the tuber of dwarf lilyturf, shizandra berry, monkshood, the root of bidentate achyranthes in an authentic gas soup formula ratio, except cultivated land, all the other medicinal material dryings, pulverize;
2) by cultivated land segment, with step 1) medicinal material after pulverizing mixes, extracts medicinal material 1-3 time with ultrasonic extraction, coarse filtration after 7-9 times of water extraction 25-35min at every turn, centrifugation after hold over night, gets supernatant liquor;
3) by step 2) in supernatant concentration to relative density be 1.15-1.2, being adjusted to liquid ethanol content with ethanol is 75-85%, leave standstill centrifugation, get lower sediment with obtaining Crude polysaccharides A in 55-65 DEG C of vacuum-drying after washing with alcohol;
4) getting step 3) Crude polysaccharides A is dissolved in water, and shakes up, and acid for adjusting pH, to 5-6, adds papoid, at 45-55 DEG C, enzymolysis 4-6h, deactivation, the centrifugal precipitation of removing, gets subnatant and obtains Crude polysaccharides enzymolysis solution B;
5) to step 4) add chloroform-butanol solution of its volume 1/4-1/6 in Crude polysaccharides enzymolysis solution B, concussion, hold over night, collects upper strata aqueous phase solution, obtains polysaccharide soln B;
6) get step 5) polysaccharide soln A, add water and be mixed with the solution that concentration is 3-5mg/ml, regulate the pH to 4.5-5.5 of polysaccharide soln, add macroporous resin to stir 1-3h under 45-55 DEG C of water-bath, the Crude polysaccharides B of filtration;
7) get step 6) Crude polysaccharides B adds water and makes the solution that concentration is 0.15-0.25mg/mL, carries out ultrafiltration, collects trapped fluid, concentrated after 55-65 DEG C of vacuum-drying, obtain smart polysaccharide.
Further, the method for the industrial extraction and purification of an above-mentioned authentic gas soup polysaccharide, step 1) described drying temperature is 60 DEG C.
Further, the method for the industrial extraction and purification of an above-mentioned authentic gas soup polysaccharide, step 2) described ultrasound condition stops 2s, power 800w for super 2s at room temperature.
The method of the industrial extraction and purification of an above-mentioned authentic gas soup polysaccharide, step 2), step 3) and step 4) described centrifugal condition is 5000rpm × 10min.
Further, the method for the industrial extraction and purification of an above-mentioned authentic gas soup polysaccharide, step 3) described supernatant concentration is to be 1.15-1.2 at 60 DEG C of rotary evaporations to relative density of medicine liquid.
Further, the method for the industrial extraction and purification of an above-mentioned authentic gas soup polysaccharide, step 4) described acid is Glacial acetic acid.
Further, the method for the industrial extraction and purification of an above-mentioned authentic gas soup polysaccharide, gets step 2) Crude polysaccharides A: water: the mass ratio of papoid is 1:50:1.5.
Further, the method for the industrial extraction and purification of an above-mentioned authentic gas soup polysaccharide, step 4) described deactivation is at boiling water bath deactivation 5min.
Further, the method of the industrial extraction and purification of an above-mentioned authentic gas soup polysaccharide, step 7) described ultrafiltration charging flow velocity is 10-14mL/min, and pressure is 0.08-012MPa, and the volume ratio that makes transit dose and interception is 1:1, in the time that solution all enters ultrafiltration post, be designated as ultrafiltration once, will in trapped fluid, add 200mL distilled water, continue ultrafiltration, circulate after 25 times, collect trapped fluid.
Compared with prior art, technical scheme provided by the invention: simple to operate, polysaccharide content is high, and purity is high, and be easy to suitability for industrialized production, by deproteinated, after uf processing, polysaccharide content all rises higher.Although itself does not improve the polysaccharide content in Crude polysaccharides deproteinated processing, this step is absolutely necessary for the treating process of whole polysaccharide.
An authentic gas soup has more significant clinical efficacy, and it is carried out to deep Study on mechanism, illustrates its scientific meaning, has important clinical meaning.Particularly, for the basic substance of its effect, be necessary to further investigate.In an authentic gas soup, multiple medicinal material contains polyose composition, and modern study also proves, polyose composition has multiple physiologically active, as improved immunizing power, antiviral etc.
Embodiment
Below in conjunction with embodiment, claim of the present invention is done to further restriction, the amendment of anyone limited number of time making within the scope of the claims in the present invention is still within the claims in the present invention protection domain.
Embodiment 1
The method of the industrial extraction and purification of an authentic gas soup polysaccharide provided by the invention, comprises the steps: successively
1) extract
Take pharmaceutical decocting piece in an authentic gas soup formula ratio, totally 6 doses, except cultivated land, 60 DEG C of all the other medicinal materials dry, pulverize as coarse particles.Cultivated land is cut into segment.Adopt ultrasonic extraction to extract medicinal material 2 times, each 8 times of water extraction 30min (ultrasound condition: super 2s stops 2s, power 800w, room temperature).United extraction liquid, by nylon wire (200 order) coarse filtration, centrifugal treating after hold over night (5000rpm × 10min), obtain supernatant liquor, 60 DEG C of rotary evaporation to relative densities of liquid are 1.15-1.2, and being adjusted to liquid ethanol content with ethanol is 80%, leave standstill 24h, the centrifugal 10min of 5000rpm, precipitation obtains Crude polysaccharides A in 60 DEG C of vacuum-dryings by 80% washing with alcohol after once.
2) deproteinated
3) getting step 1) Crude polysaccharides A2.0g is in triangular flask, and the 100mL that adds water dissolves, and shakes up, and Glacial acetic acid regulates pH to 5.5, adds papoid 3g, and 50 DEG C, enzymolysis 5h, boiling water bath deactivation 5min, the centrifugal precipitation of removing of 5000rpm × 10min.Obtain Crude polysaccharides enzymolysis solution.Adopt Sevag method, in above-mentioned enzymolysis solution, add the chloroform-butanol solution of its volume 1/5, concussion 30min, hold over night, collects upper strata aqueous phase solution, in triplicate, obtains deproteinated Crude polysaccharides B.
3) depigmentation
4) get step 2) deproteinated Crude polysaccharides B, add water and be mixed with the solution that concentration is about 4mg/ml, regulate the pH to 5.0 of polysaccharide soln with Glacial acetic acid, add 2g macroporous resin AB-8 (weight percent is 5%), under 50 DEG C of water-baths, stir 2h, filter deproteinated depigmentation Crude polysaccharides B.
4) ultrafiltration
Get step 3) deproteinated depigmentation Crude polysaccharides B80mg, add water and make the solution that concentration is about 0.2mg/mL, carry out ultrafiltration, charging flow velocity is 12mL/min, and adjusting pressure is 0.1MPa, and the volume ratio that makes transit dose and interception is 1:1, in the time that solution all enters ultrafiltration post, be designated as ultrafiltration once, will in trapped fluid, add 200mL distilled water, continue ultrafiltration, circulate after 25 times, collect trapped fluid, 60 DEG C of Rotary Evaporators are concentrated on a small quantity, after 60 DEG C of vacuum-drying, survey polysaccharide content.
For higher explanation the present invention advantage place compared with prior art, provide research method of the present invention below:
1 instrument and reagent
Ultrasonic circulating extracting machine (TGCX2-2B, great auspicious grand), ultraviolet-visible spectrophotometer (TU-U901, Beijing Puxi General Instrument Co., Ltd), thermostat water bath (HWS-24, Shanghai Yiheng Scientific Instruments Co., Ltd), electronic balance (XS105DU, plum Teller-Tuo benefit), vacuum drying oven (DZF-6050, Shanghai Yiheng Scientific Instruments Co., Ltd), Rotary Evaporators (R-3, BUCHI).
Radix Rehmanniae Preparata, the bighead atractylodes rhizome, Radix Codonopsis, the tuber of dwarf lilyturf, shizandra berry, monkshood, the root of bidentate achyranthes (prepared slices of Chinese crude drugs factory of medicinal material company of Guangdong Province), macroporous resin (AB-8, Rui Ou bio tech ltd, Tianjin), glucose (sigama), phenol, the vitriol oil, hydrogen peroxide, gacs etc. are domestic analytical pure.Bovine serum albumin (purity >=96%, sigama), Xylene Brilliant Cyanine G, chloroform, propyl carbinols etc. are domestic analytical pure.
2. method and result
2.1 determination of polysaccharide
Adopt phenolsulfuric acid method.
2.1.1 the preparation of reference substance stock solution
Precision takes in glucose reference substance 20.92mg to 100mL measuring bottle, is dissolved in water, and shakes up, and is settled to scale, and obtaining concentration is the glucose stock solution of 0.2092mg/mL.
2.1.2 the drafting of typical curve
Precision measures reference substance stock solution 0.1,0.2,0.4,0.6,0.8,1.0mL in 10mL tool plug test tube respectively, mends and adds water to 2.0mL, shakes up.Add 5% phenol solution 1mL, shake up, add rapidly vitriol oil 5mL, put and in boiling water bath, be incubated 15min, take out to put and in ice bath, be cooled to room temperature.Separately getting distilled water 2.0mL puts in tool plug test tube, add the aforesaid method processing of corresponding reagent, prepare blank solution, measure absorbancy at 490nm place, by absorbancy, glucose quality is carried out to linear regression, obtain regression equation: Y=7.404X+0.022 (r=0.9997)
2.1.3 the assay of sample
Get the about 10mg of trial-product, accurately weighedly in 50mL measuring bottle, be dissolved in water, shake up, and be settled to scale.Precision measures 0.4mL in 10mL tool plug test tube, mends and adds water to 2.0mL, by method processing under 2.2.2 item, and measures.
The mensuration of 2.2 protein contents
Adopt Xylene Brilliant Cyanine G method.
2.2.1 the preparation of Xylene Brilliant Cyanine G G-250 solution
Take Xylene Brilliant Cyanine G G-25025mg in 250mL measuring bottle, add 12.5mL95% dissolve with ethanol, shake up, add 25mL85%H
3pO
4, after shaking up, adding and be diluted to scale, 5000rpm × 10min centrifuging and taking supernatant liquor is for subsequent use.
2.2.2 the preparation of reference substance stock solution
Precision takes in bovine serum albumin 10.51mg to 50mL measuring bottle, is dissolved in water, and shakes up, and is settled to scale, and obtaining concentration is the bovine serum albumin stock solution of 0.2102mg/mL.
2.2.3 the drafting of typical curve
Precision measures reference substance stock solution 0.0,0.1,0.2,0.4,0.6,0.8,1.0mL in 25mL tool plug test tube respectively, mends and adds water to 2.0mL, shakes up.Add Xylene Brilliant Cyanine G G-250 solution 10mL, shake up, room temperature is placed 30min, taking distilled water as blank, measure absorbancy at 595nm wavelength place, by absorbancy, the quality of bovine serum albumin is carried out to linear regression, obtain regression equation: Y=2.298X+1.403 (r=0.993)
2.2.4 the preparation of trial-product and the mensuration of content
Get the about 10mg of Crude polysaccharides, accurately weighedly in 50mL measuring bottle, be dissolved in water, shake up, and be settled to scale.Precision measures 2mL in 10mL tool plug test tube, by method processing under 2.2.3 item, measures and calculate protein content.
2.3 extract
Take pharmaceutical decocting piece in an authentic gas soup formula ratio, totally 6 doses, except cultivated land, 60 DEG C of all the other medicinal materials dry, pulverize as coarse particles.Cultivated land is cut into segment.Adopt ultrasonic extraction to extract medicinal material 2 times, each 8 times of water extraction 30min (ultrasound condition: super 2s stops 2s, power 800w, room temperature).United extraction liquid, by nylon wire (200 order) coarse filtration, centrifugal treating after hold over night (5000rpm × 10min), obtain supernatant liquor, 60 DEG C of rotary evaporation to relative densities of liquid are 1.15-1.2, and being adjusted to liquid ethanol content with ethanol is 80%, leave standstill 24h, the centrifugal 10min of 5000rpm, precipitation obtains Crude polysaccharides A in 60 DEG C of vacuum-dryings by 80% washing with alcohol after once.Survey polysaccharide, protein content by 2.1,2.2 lower methods respectively.The results are shown in Table 1
2.4 deproteinated
Get Crude polysaccharides A2.0g in triangular flask, the 100mL that adds water dissolves, and shakes up, and Glacial acetic acid regulates pH to 5.5, adds papoid 3g, and 50 DEG C, enzymolysis 5h, boiling water bath deactivation 5min, the centrifugal precipitation of removing of 5000rpm × 10min.Obtain Crude polysaccharides enzymolysis solution.Adopt Sevag method, in above-mentioned enzymolysis solution, add the chloroform-butanol solution of its volume 1/5, concussion 30min, hold over night, collects upper strata aqueous phase solution, in triplicate, surveys polysaccharide and protein content respectively according to 2.1,2.2 lower methods.The results are shown in Table 1.
As shown in Table 1, an authentic gas soup Crude polysaccharides is carried out after deproteinated processing, protein content drops to 4.61 ± 0.09% by 9.83 ± 0.23%, but the content of polysaccharide drops to 37.00 ± 0.74% by 44.18 ± 1.02% simultaneously, result shows, deproteinated process, and polysaccharide has loss.
The polysaccharide content of table 1 deproteinated on Crude polysaccharides and the impact of protein content (n=3)
| Numbering | Polysaccharide content (%) | Protein content (%) |
| Crude polysaccharides A | 44.18±1.02 | 9.83±0.23 |
| Deproteinated Crude polysaccharides B | 37.00±0.74 | 4.61±0.09 |
The investigation of 2.5 depigmentation methods
2.5.1 to get Crude polysaccharides appropriate for pigment detection method, be mixed with the solution that concentration is 2mg/mL, solution colour is brown color, from complementary color principle, when the light time of certain color in the absorption visible ray of certain matter selective of solution, solution will present corresponding complementary color, because Radix Ophiopogonis polysaccharide solution is orange-yellow, proving has maximum absorption to its complementary light blueness, therefore select 450nm as detecting wavelength, measure the absorbancy of polysaccharide soln depigmentation front and back, calculate pigment decreasing ratio according to following formula:
Pigment decreasing ratio %=× 100%
2.5.2 an authentic gas soup polysaccharide depigmentation method relatively to get Crude polysaccharides appropriate, add water and be mixed with the solution that concentration is about 4mg/ml, be divided into 12 parts, get 9 parts of comparisons for depigmentation method, 3 every group parallel.
(1) macroporous resin AB-8 static adsorptive method decolouring: the pH to 5.0 that regulates polysaccharide soln with Glacial acetic acid, add 2g macroporous resin AB-8 (weight percent is 5%), under 50 DEG C of water-baths, stir 2h, after filtration, be taken at 450nm wavelength place and measure light absorption value A, calculate pigment decreasing ratio.Sampling 20mL, measures polysaccharide content by 2.2 lower methods after 60 DEG C of drying under reduced pressure.The results are shown in Table 2, the average percent of decolourization of this method is 48.32 ± 1.05%.
(2) activated carbon method decolouring: the pH to 5.0 that regulates polysaccharide soln with Glacial acetic acid, add 1.2g gac (weight percent is 3%), under 50 DEG C of water-baths, stir 1h, after filtration, measure light absorption value A at 450nm wavelength place, calculate pigment decreasing ratio.Sampling 20mL, measures polysaccharide content by 2.2 lower methods after 60 DEG C of drying under reduced pressure.The results are shown in Table 2, the average percent of decolourization of this method is 68.49 ± 1.02%.
(3) H
2o
2method decolouring: regulate the pH to 8.0 of polysaccharide soln with NaOH solution, drip H
2o
2(volume percent is 20%) stirs 3h under 50 DEG C of water-baths, measures light absorption value A after filtration at 450nm wavelength place, calculates pigment decreasing ratio.Sampling 20mL, measures polysaccharide content by 2.2 lower methods after 60 DEG C of drying under reduced pressure.The results are shown in Table 2, the average percent of decolourization of this method is 68.98 ± 0.69%.
As shown in Table 2, H
2o
2method and activated carbon method pigment decreasing ratio are high compared with Amberlyst process, but the destruction of polysaccharide is many simultaneously, and polysaccharide content declines more, therefore select Amberlyst process decolouring.
The authentic gas soup polysaccharide depigmentation method comparison (n=3) of table 2
| Decoloring method | Polysaccharide content (%) | A 450nm | Pigment decreasing ratio (%) |
| No bleaching | 37.00±0.94 | 1.190±0.005 | -- |
| Macroporous resin AB-8 | 39.63±1.08 | 0.615±0.013 | 48.32±1.05 |
?
| Gac | 32.19±0.86 | 0.375±0.012 | 68.49±1.02 |
| H 2O 2 | 33.98±1.19 | 0.205±0.010 | 68.98±0.69 |
Refining of 2.6 polysaccharide
Adopt ultrafiltration process to refine Crude polysaccharides A.
2.6.1 ultrafiltration
Get respectively Crude polysaccharides A80mg, add water and make the solution that concentration is about 0.2mg/mL, carry out ultrafiltration, charging flow velocity is 12mL/min, and adjusting pressure is 0.1MPa, and the volume ratio that makes transit dose and interception is 1:1, in the time that solution all enters ultrafiltration post, be designated as ultrafiltration once, will in trapped fluid, add 200mL distilled water, continue ultrafiltration, circulate after 25 times, collect trapped fluid, 60 DEG C of Rotary Evaporators are concentrated on a small quantity, after 60 DEG C of vacuum-drying, survey respectively polysaccharide content 51.16 ± 0.94%.
Determining of 2.7 polysaccharide purification techniques
2.7.1 the impact of deproteinated on purifying process
By ultrafiltration process, to Crude polysaccharides A, deproteinated Crude polysaccharides B refines.Measure polysaccharide content, the results are shown in Table 3.
In table 3, contrast technique 1 and 3 is known, refines by ultrafiltration process, and the polysaccharide content of Crude polysaccharides A has risen about 7%; Contrast technique 2 and 4 is known, and the Crude polysaccharides B content after deproteinated has risen 29.4%.
The impact (n=3) of table 3 deproteinated on an authentic gas soup polysaccharide purification technique
| Numbering | Preparation technology | Polysaccharide content (%) |
| 1 | Water extraction → alcohol precipitation | 44.18±1.02 |
| 2 | Water extraction → alcohol precipitation → deproteinated | 37.00±0.74 |
| 3 | Water extraction → alcohol precipitation → ultrafiltration | 51.16±0.94 |
| 4 | Water extraction → alcohol precipitation → deproteinated → ultrafiltration | 66.40±0.71 |
2.7.2 the impact of depigmentation on purifying process
Adopt ultrafiltration process to refine the Crude polysaccharides of deproteinated Crude polysaccharides B and (deproteinated+depigmentation) processing, measure polysaccharide content, the results are shown in Table 4.
In table 4, contrast technique 1 and 2 is known, and after depigmentation is processed, the polysaccharide content of an authentic gas soup has raising by a small margin, and contrast technique 2 and 3 is known, and the polysaccharide after depigmentation is again after uf processing, and polysaccharide content improves more, reaches 81%.
The impact (n=3) of table 4 depigmentation on an authentic gas soup polysaccharide purification technique
| Numbering | Preparation technology | Polysaccharide content (%) |
| 1 | Water extraction → alcohol precipitation → deproteinated | 37.00±0.74 |
?
| 2 | Water extraction → alcohol precipitation → deproteinated → depigmentation | 39.63±1.08 |
| 3 | Water extraction → alcohol precipitation → deproteinated → ultrafiltration | 66.40±0.71 |
| 4 | Water extraction → alcohol precipitation → deproteinated → depigmentation → ultrafiltration | 81.14±0.75 |
2.7.3 determining of purifying process
According to table 3,4 results, determine that the extraction and purification process of an authentic gas sugar soup polysaccharide is: water extraction → alcohol precipitation → deproteinated → depigmentation → ultrafiltration, under these processing condition, can prepare high purity polysaccharide.
Three kinds of depigmentation methods of the authentic gas soup polysaccharide that this institute adopts, are gained optimum process condition after optimizing.Visible by comprehensive relatively depigmentation rate and polysaccharide content, the decolorizing effect of gac and hydrogen peroxide is better, but the adsorption of gac is non-selectivity absorption, therefore polysaccharide can lose, and the strong oxidizing property of hydrogen peroxide may cause the change of polysaccharide structures, therefore after these two kinds of decoloring methods are processed, polysaccharide content also declines to some extent.The decreasing ratio of macroporous resin AB-8 static adsorptive method pigment is slightly lower than other two kinds of methods, but polysaccharide content increases after this method is processed, and this method is simple to operate, considers and thinks that the pigment that is applicable to an authentic gas soup Crude polysaccharides removes.In addition, table 4 result shows: although depigmentation is less to the content influence of polysaccharide, and first depigmentation, then pass through ultrafiltration, and polysaccharide content finally can reach more than 80%, and depigmentation is very important step in an authentic gas soup polysaccharide purification technique as can be seen here.
As shown in Table 1, an authentic gas soup is carried out after deproteinated processing, when albumen effective elimination, the content of polysaccharide also declines to some extent.But table 3 data show, deproteinated process after polysaccharide content suppression ratio more, all rise higher by the polysaccharide content after uf processing.Presentation of results, although deproteinated processing itself does not improve the polysaccharide content in Crude polysaccharides, this step is absolutely necessary for the treating process of whole polysaccharide.
As shown in Table 4, Crude polysaccharides A, deproteinated Crude polysaccharides B is through ultrafiltration process, and after ultrafiltration process is refining, the content of polysaccharide is improved, and is easy to industrialization operation.
An authentic gas soup has more significant clinical efficacy, and it is carried out to deep Study on mechanism, illustrates its scientific meaning, has important clinical meaning.Particularly, for the basic substance of its effect, be necessary to further investigate.In an authentic gas soup, multiple medicinal material contains polyose composition, and modern study also proves, polyose composition has multiple physiologically active, as improved immunizing power, antiviral etc.
Claims (9)
1. a method for the industrial extraction and purification of an authentic gas soup polysaccharide, is characterized in that, comprises the steps: successively
1) take Radix Rehmanniae Preparata, the bighead atractylodes rhizome, ginseng, the tuber of dwarf lilyturf, shizandra berry, monkshood, the root of bidentate achyranthes in an authentic gas soup formula ratio, except cultivated land, all the other medicinal material dryings, pulverize;
2) by cultivated land segment, with step 1) medicinal material after pulverizing mixes, extracts medicinal material 1-3 time with ultrasonic extraction, coarse filtration after 7-9 times of water extraction 25-35min at every turn, centrifugation after hold over night, gets supernatant liquor;
3) by step 2) in supernatant concentration to relative density be 1.15-1.2, being adjusted to liquid ethanol content with ethanol is 75-85%, leave standstill centrifugation, get lower sediment with obtaining Crude polysaccharides A in 55-65 DEG C of vacuum-drying after washing with alcohol;
4) getting step 3) Crude polysaccharides A is dissolved in water, and shakes up, and acid for adjusting pH, to 5-6, adds papoid, at 45-55 DEG C, enzymolysis 4-6h, deactivation, the centrifugal precipitation of removing, gets subnatant and obtains Crude polysaccharides enzymolysis solution B;
5) to step 4) add chloroform-butanol solution of its volume 1/4-1/6 in Crude polysaccharides enzymolysis solution B, concussion, hold over night, collects upper strata aqueous phase solution, obtains polysaccharide soln B;
6) get step 5) polysaccharide soln A, add water and be mixed with the solution that concentration is 3-5mg/ml, regulate the pH to 4.5-5.5 of polysaccharide soln, add macroporous resin to stir 1-3h under 45-55 DEG C of water-bath, the Crude polysaccharides B of filtration;
7) get step 6) Crude polysaccharides B adds water and makes the solution that concentration is 0.15-0.25mg/mL, carries out ultrafiltration, collects trapped fluid, concentrated after 55-65 DEG C of vacuum-drying, obtain smart polysaccharide.
2. according to the method for the industrial extraction and purification of an authentic gas soup polysaccharide claimed in claim 1, it is characterized in that step 1) described drying temperature is 60 DEG C.
3. the method for the industrial extraction and purification of an authentic gas soup polysaccharide according to claim 1, is characterized in that step 2) described ultrasound condition stops 2s, power 800w for super 2s at room temperature.
4. the method for the industrial extraction and purification of an authentic gas soup polysaccharide according to claim 1, is characterized in that step 2), step 3) and step 4) described centrifugal condition is 5000rpm × 10min.
5. the method for the industrial extraction and purification of an authentic gas soup polysaccharide according to claim 1, is characterized in that step 3) described supernatant concentration is to be 1.15-1.2 at 60 DEG C of rotary evaporations to relative density of medicine liquid.
6. the method for the industrial extraction and purification of an authentic gas soup polysaccharide according to claim 1, is characterized in that step 4) described acid is Glacial acetic acid.
7. the method for the industrial extraction and purification of an authentic gas soup polysaccharide according to claim 1, is characterized in that, gets step 2) Crude polysaccharides A: water: the mass ratio of papoid is 1:50:1.5.
8. the method for the industrial extraction and purification of an authentic gas soup polysaccharide according to claim 1, is characterized in that step 4) described deactivation is at boiling water bath deactivation 5min.
9. the method for the industrial extraction and purification of an authentic gas soup polysaccharide according to claim 1, it is characterized in that, step 7) described ultrafiltration charging flow velocity is 10-14mL/min, and pressure is 0.08-012MPa, and the volume ratio that makes transit dose and interception is 1:1, in the time that solution all enters ultrafiltration post, be designated as ultrafiltration once, will in trapped fluid, add 200mL distilled water, continue ultrafiltration, circulate after 25 times, collect trapped fluid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410188735.1A CN104031155B (en) | 2014-05-06 | 2014-05-06 | The method of the industrial extraction and purification of an authentic gas soup polysaccharide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410188735.1A CN104031155B (en) | 2014-05-06 | 2014-05-06 | The method of the industrial extraction and purification of an authentic gas soup polysaccharide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104031155A true CN104031155A (en) | 2014-09-10 |
| CN104031155B CN104031155B (en) | 2016-06-01 |
Family
ID=51462168
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410188735.1A Active CN104031155B (en) | 2014-05-06 | 2014-05-06 | The method of the industrial extraction and purification of an authentic gas soup polysaccharide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104031155B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106749746A (en) * | 2017-02-27 | 2017-05-31 | 云南省热带作物科学研究所 | It is a kind of to improve the method and extract obtained and application for obtaining the beautiful polysaccharide of Noni fruit promise |
| CN115232225A (en) * | 2022-08-17 | 2022-10-25 | 漳州卫生职业学院 | Prepared rehmannia root homogeneous polysaccharide and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009125964A2 (en) * | 2008-04-07 | 2009-10-15 | Korea Institute Of Radiological & Medical Sciences | Composition comprising polysaccharide extracted from panax ginseng preventing and treating liver diseases |
| CN101838336A (en) * | 2009-03-20 | 2010-09-22 | 华中科技大学 | Hubei dwarf lilyturf tuber hetetopolysaccharide for curing type II diabete and preparation method thereof |
| CN103301290A (en) * | 2013-06-25 | 2013-09-18 | 广西医科大学 | Compound longan polysaccharide immune foundation-strengthening prescription and preparation method thereof |
-
2014
- 2014-05-06 CN CN201410188735.1A patent/CN104031155B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009125964A2 (en) * | 2008-04-07 | 2009-10-15 | Korea Institute Of Radiological & Medical Sciences | Composition comprising polysaccharide extracted from panax ginseng preventing and treating liver diseases |
| CN101838336A (en) * | 2009-03-20 | 2010-09-22 | 华中科技大学 | Hubei dwarf lilyturf tuber hetetopolysaccharide for curing type II diabete and preparation method thereof |
| CN103301290A (en) * | 2013-06-25 | 2013-09-18 | 广西医科大学 | Compound longan polysaccharide immune foundation-strengthening prescription and preparation method thereof |
Non-Patent Citations (3)
| Title |
|---|
| 李赞阳 等: "大孔树脂分离纯化中药复方多糖的工艺研究", 《石河子大学学报（自然科学版）》 * |
| 陈虹: "全真一气汤总多糖的提取与质量控制初步研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
| 靳学远 等: "《天然产物降血糖功能性成分研究》", 30 June 2009, 上海交通大学出版社 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106749746A (en) * | 2017-02-27 | 2017-05-31 | 云南省热带作物科学研究所 | It is a kind of to improve the method and extract obtained and application for obtaining the beautiful polysaccharide of Noni fruit promise |
| CN106749746B (en) * | 2017-02-27 | 2019-06-14 | 云南省热带作物科学研究所 | It is a kind of to improve the method for obtaining the beautiful polysaccharide of Noni fruit promise and extract obtained and application |
| CN115232225A (en) * | 2022-08-17 | 2022-10-25 | 漳州卫生职业学院 | Prepared rehmannia root homogeneous polysaccharide and preparation method and application thereof |
| CN115232225B (en) * | 2022-08-17 | 2023-04-14 | 漳州卫生职业学院 | Prepared rehmannia root homogeneous polysaccharide and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104031155B (en) | 2016-06-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102146142B (en) | Method for preparing astragalus polysaccharides | |
| CN102417544A (en) | Ziyang selenium-rich green tea selenium-containing polysaccharide, and preparation method and application thereof | |
| CN110551230B (en) | Preparation method of astragalus polysaccharide | |
| CN109400741B (en) | Separation and purification method of ganoderma lucidum spore polysaccharide | |
| CN106214845A (en) | A kind of pharmaceutical composition with health care and preparation method thereof | |
| CN105663444A (en) | Compound immunity-enhancing and aging-resisting agent and preparation method thereof | |
| CN104031155B (en) | The method of the industrial extraction and purification of an authentic gas soup polysaccharide | |
| CN105399795B (en) | Method for extracting astragaloside from radix astragali | |
| CN107722131B (en) | Total ganoderma lucidum spore powder refined polysaccharide with significant auxiliary antitumor activity and preparation method and application thereof | |
| CN103113489B (en) | Method of purifying polysaccharide of Xinjiang jun dates | |
| CN103505486A (en) | Lucid ganoderma-American ginseng granule and preparation method thereof | |
| CN105777921A (en) | Astragalus polysaccharide extraction process and application thereof | |
| CN103992415B (en) | The method of the extraction purification of an authentic gas soup polysaccharide | |
| CN107245113A (en) | Corn silk polysaccharide extract with antitumaous effect and preparation method thereof | |
| CN1884571B (en) | Process for extracting polysaccharide from lycoris plant | |
| CN109535272A (en) | A kind of extracting method of pear flesh selenium polysaccharide | |
| CN105147716A (en) | Newcastle disease vaccine Yupingfeng polysaccharide immunopotentiator | |
| CN109354601A (en) | The extracting method of selenoprotein in a kind of pear fruit | |
| CN104945532A (en) | Preparing method for gynura divaricata polysaccharide | |
| CN101643515A (en) | Method for separating and purifying lentinan for injection | |
| CN103554289A (en) | Rhizoma atractylodis sinensis polysaccharide and extraction method and applications thereof in preparing anti-tumor medicaments | |
| CN105560173B (en) | A kind of high stability astragalus injection and preparation method thereof | |
| CN108164571A (en) | A kind of mung bean alpha-galactooligosaccharide extract and its extracting method and skin care application | |
| CN101537056B (en) | Method for preparing Chinese medicinal preparation for treating external respiration infection of children | |
| CN100554326C (en) | A kind of sealwort immune polysaccharide, its composition and its purposes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |