CN104031155B - The method of the industrial extraction and purification of an authentic gas soup polysaccharide - Google Patents
The method of the industrial extraction and purification of an authentic gas soup polysaccharide Download PDFInfo
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Abstract
The present invention discloses the method for the industrial extraction and purification of an a kind of authentic gas soup polysaccharide, belongs to traditional Chinese medicine extraction technology, it is intended to provide one method simple, the extracting and purifying method that polysaccharide content is high; The method comprises and takes each component, and all the other medicinal material dryings except cultivated land are pulverized; By cultivated land segment, extracting with ultrasonic extraction, supernatant liquor is got in centrifugation; By supernatant concentration, being adjusted to liquid ethanol content is 75-85%, leave standstill centrifugation, get lower sediment dry Crude polysaccharides A; Getting Crude polysaccharides A to be dissolved in water, shake even, acid for adjusting pH, to 5-6, adds papoid, enzymolysis, and the centrifugal removing precipitation of deactivation, gets subnatant and obtain Crude polysaccharides enzymolysis solution A; In Crude polysaccharides enzymolysis solution A, add chloroform-butanol solution, concussion, hold over night, collect upper strata aqueous phase solution, obtain polysaccharide soln A; Add water and it is mixed with solution, regulate the pH to 4.5-5.5 of polysaccharide soln, add macroporous resin adsorption, filter to obtain Crude polysaccharides, by solution ultrafiltration; Refining polysaccharide can be obtained.
Description
Technical field
The present invention relates to the extracting and purifying method of a kind of polysaccharide, particularly relate to the method for the industrial extraction and purification of an a kind of green authentic gas soup polysaccharide; Belong to technical field of extraction of Chinese traditional medicine.
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 ". It is made up of Radix Rehmanniae Preparata, the bighead atractylodes rhizome, ginseng, the tuber of dwarf lilyturf, shizandra berry, monkshood, the root of bidentate achyranthes. Clinical be extensively used for the treatment of deficient property disease in various old age, such as gastrointestinal illness, chronic obstructive pulmonary disease, lung cancer, etc. dialectical genus negative and positive of qi and blood all empty immunologic hypofunction persons, evident in efficacy. Modern study shows, polysaccharide has immunity moderation, and reducing blood-fat is antitumor, anti-inflammation, anti-ageing physiological function of waiting for a long time.
The current research report of the extraction about an authentic gas soup polysaccharide is seldom, an authentic gas soup polysaccharide has been carried out discussion to a certain degree by this seminar in recent years, find that its main pharmacological action is strengthening immunity, the non-specific immune function of normal mouse and endoxan immunosuppressed mice is had enhancement in various degree, the humoral immune function that endoxan suppresses can be improved, there is intestinal mucosa immunoregulation effect, its dominant mechanism is for raising immunosuppressed mice blood serum IL-6 and TNF-�� level, immune stimulatory suppresses the cytokine secretion of mouse body, thus reach the effect of immunity moderation function.
Although Chen Hong provides " extraction of an authentic gas soup total polysaccharides and quality control preliminary study " in Master's thesis, but the polysaccharide content of gained is not high, 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) taking 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) pulverize after medicinal material mix, by ultrasonic extraction extraction medicinal material 1-3 time, coarse filtration after each 7-9 times of water extraction 25-35min, 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, obtain Crude polysaccharides A in 55-65 DEG C of vacuum-drying after getting lower sediment washing with alcohol;
4) step 3 is got) Crude polysaccharides A is dissolved in water, and shakes even, and acid for adjusting pH, to 5-6, adds papoid, at 45-55 DEG C, enzymolysis 4-6h, deactivation, centrifugal removing precipitates, and gets subnatant and obtains Crude polysaccharides enzymolysis solution B;
5) to step 4) Crude polysaccharides enzymolysis solution B adds the chloroform-butanol solution of its volume 1/4-1/6, concussion, hold over night, collects upper strata aqueous phase solution, obtains polysaccharide soln B;
6) step 5 is got) polysaccharide soln A, add water and it is mixed with the solution that concentration is 3-5mg/ml, regulate the pH to 4.5-5.5 of polysaccharide soln, add macroporous resin and stir 1-3h under 45-55 DEG C of water-bath, the Crude polysaccharides B of filtration;
7) step 6 is got) Crude polysaccharides B adds water and makes the solution that concentration is 0.15-0.25mg/mL, carries out ultrafiltration, collects and retains liquid, 55-65 DEG C of vacuum-drying after concentrated, obtains essence polysaccharide.
Further, the method for the industrial extraction and purification of an above-mentioned authentic gas soup polysaccharide, step 1) described in drying temperature be 60 DEG C.
Further, the method for the industrial extraction and purification of an above-mentioned authentic gas soup polysaccharide, step 2) described in ultrasound condition stop 2s, power 800w at room temperature super 2s.
The method of the industrial extraction and purification of an above-mentioned authentic gas soup polysaccharide, step 2), step 3) and step 4) described in centrifugal condition be 5000rpm �� 10min.
Further, the method for the industrial extraction and purification of an above-mentioned authentic gas soup polysaccharide, step 3) described in supernatant concentration 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 in acid be 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 in deactivation be 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 in ultrafiltration charging flow velocity be 10-14mL/min, pressure is 0.08-012MPa so that be 1:1 through amount with the volume ratio of interception, when solution all enters ultrafiltration post, it is designated as ultrafiltration once, will retain in liquid and add 200mL distilled water, continue ultrafiltration, circulate after 25 times, collect and retain liquid.
Compared with prior art, technical scheme provided by the invention: simple to operate, polysaccharide content height, purity height, and it is easy to suitability for industrialized production, by deproteinated, after uf processing, polysaccharide content all rises higher. Although deproteinated process itself does not improve the polysaccharide content in Crude polysaccharides, but this step is essential for the treating process of whole polysaccharide.
An authentic gas soup has more significant clinical efficacy, it is carried out deep Study on mechanism, illustrates its scientific meaning, have important clinical meaning. Particularly for the basic substance of its effect, it is 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, the claim of the present invention being done further restriction, the amendment of anyone limited time made 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
Taking pharmaceutical decocting piece, totally 6 doses in an authentic gas soup formula ratio, except cultivated land, the 60 DEG C of dryings of all the other medicinal materials, pulverize as coarse particles. Cultivated land is cut into segment. Ultrasonic extraction is adopted 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 (5000rpm �� 10min) after hold over night, obtain supernatant liquor, 60 DEG C of rotary evaporations are 1.15-1.2 to the relative density of liquid, and being adjusted to liquid ethanol content with ethanol is 80%, leave standstill 24h, the centrifugal 10min of 5000rpm, precipitate with 80% washing with alcohol once after obtain Crude polysaccharides A in 60 DEG C of vacuum-dryings.
2) deproteinated
3) step 1 is got) Crude polysaccharides A2.0g is in triangular flask, and the 100mL that adds water dissolves, and shakes even, and Glacial acetic acid regulates pH to 5.5, adds papoid 3g, 50 DEG C, enzymolysis 5h, the centrifugal removing precipitation of boiling water bath deactivation 5min, 5000rpm �� 10min. Obtain Crude polysaccharides enzymolysis solution. Adopting Sevag method, add the chloroform-butanol solution of its volume 1/5 in above-mentioned enzymolysis solution, concussion 30min, hold over night, collects upper strata aqueous phase solution, repeats three times, obtain deproteinated Crude polysaccharides B.
3) depigmentation
4) step 2 is got) deproteinated Crude polysaccharides B, add water and it is mixed with the solution that concentration is about 4mg/ml, the pH to 5.0 of polysaccharide soln is regulated with Glacial acetic acid, add 2g macroporous resin AB-8 (weight percent is 5%), 2h is stirred under 50 DEG C of water-baths, 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 adjustment pressure is 0.1MPa so that the volume ratio passing through amount and interception is 1:1, when solution all enters ultrafiltration post, it is designated as ultrafiltration once, will retain in liquid and add 200mL distilled water, continue ultrafiltration, circulating after 25 times, collect and retain liquid, 60 DEG C of Rotary Evaporators concentrate on a small quantity, after 60 DEG C of vacuum-drying, survey polysaccharide content.
In order to higher explanation the present invention advantage place compared with prior art, provide the research method of the present invention below:
1 instrument and reagent
Ultrasonic and circulated extraction 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, Mei Tele-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, gac etc. are domestic analytical pure. Bovine serum albumin (purity >=96%, sigama), Xylene Brilliant Cyanine G, chloroform, propyl carbinol etc. are domestic analytical pure.
2. method and result
2.1 determination of polysaccharide
Adopt phend-sulphuric acid.
2.1.1 the preparation of reference substance stock solution
Precision takes in glucose control product 20.92mg to 100mL measuring bottle, is dissolved in water, shakes even, and be settled to scale, obtains the glucose stock solution that concentration is 0.2092mg/mL.
2.1.2 the drafting of typical curve
Respectively precision measure reference substance stock solution 0.1,0.2,0.4,0.6,0.8,1.0mL in 10mL tool plug test tube, mend and add water to 2.0mL, shake even. Adding 5% phenol solution 1mL, shake even, add rapidly vitriol oil 5mL, put and be incubated 15min in boiling water bath, taking-up is put and is cooled to room temperature in ice bath. Separately getting distilled water 2.0mL puts in tool plug test tube, add corresponding reagent aforesaid method to process, prepare blank solution, measure absorbancy at 490nm place, by absorbancy, glucose quality is carried out linear regression, obtain regression equation: Y=7.404X+0.022 (r=0.9997)
2.1.3 the assay of sample
Get trial-product and it is about 10mg, accurately weighed be dissolved in water in 50mL measuring bottle, shake even, and be settled to scale. Precision measures 0.4mL in 10mL tool plug test tube, mends and adds water to 2.0mL, by the process of method under 2.2.2 item, and measures.
The mensuration of 2.2 protein contents
Adopt Coomassie Brilliant Blue.
2.2.1 the preparation of Coomassie brilliant G-250 solution
Take Coomassie brilliant G-250 25mg in 250mL measuring bottle, add 12.5mL95% dissolve with ethanol, shake even, add 25mL85%H3PO4, shake even after, add 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 even, and is settled to scale, obtains the bovine serum albumin stock solution that concentration is 0.2102mg/mL.
2.2.3 the drafting of typical curve
Respectively precision measure reference substance stock solution 0.0,0.1,0.2,0.4,0.6,0.8,1.0mL in 25mL tool plug test tube, mend and add water to 2.0mL, shake even. Adding Coomassie brilliant G-250 solution 10mL, shake even, room temperature places 30min, take distilled water as blank, measure absorbancy at 595nm wavelength place, by absorbancy, the quality of bovine serum albumin is carried out 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 Crude polysaccharides and it is about 10mg, accurately weighed be dissolved in water in 50mL measuring bottle, shake even, and be settled to scale. Precision measures 2mL in 10mL tool plug test tube, by the process of method under 2.2.3 item, measures and calculates protein content.
2.3 extract
Taking pharmaceutical decocting piece, totally 6 doses in an authentic gas soup formula ratio, except cultivated land, the 60 DEG C of dryings of all the other medicinal materials, pulverize as coarse particles. Cultivated land is cut into segment. Ultrasonic extraction is adopted 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 (5000rpm �� 10min) after hold over night, obtain supernatant liquor, 60 DEG C of rotary evaporations are 1.15-1.2 to the relative density of liquid, and being adjusted to liquid ethanol content with ethanol is 80%, leave standstill 24h, the centrifugal 10min of 5000rpm, precipitate with 80% washing with alcohol once after obtain Crude polysaccharides A in 60 DEG C of vacuum-dryings. Under pressing 2.1,2.2 item respectively, method surveys polysaccharide, protein content. The results are shown in Table 1
2.4 deproteinated
Getting Crude polysaccharides A2.0g in triangular flask, the 100mL that adds water dissolves, and shakes even, and Glacial acetic acid regulates pH to 5.5, adds papoid 3g, 50 DEG C, enzymolysis 5h, the centrifugal removing precipitation of boiling water bath deactivation 5min, 5000rpm �� 10min. Obtain Crude polysaccharides enzymolysis solution. Adopting Sevag method, add the chloroform-butanol solution of its volume 1/5 in above-mentioned enzymolysis solution, concussion 30min, hold over night, collects upper strata aqueous phase solution, repeats three times, surveys polysaccharide and protein content respectively according to method under 2.1,2.2 item. The results are shown in Table 1.
As shown in Table 1, after an authentic gas soup Crude polysaccharides is carried out deproteinated process, 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, polysaccharide has loss.
Table 1 deproteinated is on the impact (n=3) of the polysaccharide content of Crude polysaccharides and protein content
| 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, it is mixed with the solution that concentration is 2mg/mL, solution colour is brown color, from complementary color principle, when the light of certain color in the absorption visible ray of certain matter selective of solution, solution will present corresponding complementary color, owing to Radix Ophiopogonis polysaccharide solution is orange-yellow, prove that its complementary light blueness is had maximum absorption, therefore select 450nm as determined wavelength, measure the absorbancy before and after polysaccharide soln depigmentation, according to following formulae discovery pigment removal rate:
Pigment removal rate %=�� 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, often organize 3 parallel.
(1) macroporous resin AB-8 static adsorptive method decolouring: the pH to 5.0 regulating polysaccharide soln with Glacial acetic acid, add 2g macroporous resin AB-8 (weight percent is 5%), 2h is stirred under 50 DEG C of water-baths, it is taken at 450nm wavelength place after filtration and measures light absorption value A, calculate pigment removal rate. 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 regulating polysaccharide soln with Glacial acetic acid, add 1.2g gac (weight percent is 3%), under 50 DEG C of water-baths, stir 1h, measure light absorption value A after filtration at 450nm wavelength place, calculate pigment removal rate. 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)H2O2Method is decoloured: the pH to 8.0 regulating polysaccharide soln by NaOH solution, drips and adds H2O2(volume percent is 20%), stirs 3h under 50 DEG C of water-baths, measures light absorption value A at 450nm wavelength place, calculate pigment removal rate after filtration. 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, H2O2Method and activated carbon method pigment removal rate relatively Amberlyst process height, but the destruction of polysaccharide is many simultaneously, and polysaccharide content decline is more, therefore selects Amberlyst process decolouring.
Authentic gas soup polysaccharide depigmentation Measures compare (n=3) of table 2
| Decoloring method | Polysaccharide content (%) | A450nm | Pigment removal rate (%) |
| 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 |
| H2O2 | 33.98��1.19 | 0.205��0.010 | 68.98��0.69 |
2.6 polysaccharide is refining
Ultrafiltration process is adopted to be refined by Crude polysaccharides A.
2.6.1 ultrafiltration
Get Crude polysaccharides A80mg respectively, add water and make the solution that concentration is about 0.2mg/mL, carry out ultrafiltration, charging flow velocity is 12mL/min, and adjustment pressure is 0.1MPa so that the volume ratio passing through amount and interception is 1:1, when solution all enters ultrafiltration post, it is designated as ultrafiltration once, will retain in liquid and add 200mL distilled water, continue ultrafiltration, circulating after 25 times, collect and retain liquid, 60 DEG C of Rotary Evaporators concentrate on a small quantity, after 60 DEG C of vacuum-drying, survey polysaccharide content 51.16 �� 0.94% respectively.
The determination of 2.7 polysaccharide purification techniques
2.7.1 deproteinated is on the impact of 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 it will be seen that refining by ultrafiltration process, and the polysaccharide content of Crude polysaccharides A rises about 7%; Contrast technique 2 and 4 is it will be seen that the Crude polysaccharides B content after deproteinated rises 29.4%.
Table 3 deproteinated is on the impact (n=3) of 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 depigmentation is on the impact of purifying process
Adopt ultrafiltration process to be refined by the Crude polysaccharides that deproteinated Crude polysaccharides B and (deproteinated+depigmentation) process, measure polysaccharide content, the results are shown in Table 4.
In table 4, contrast technique 1 and 2 is it will be seen that after depigmentation process, the polysaccharide content of an authentic gas soup has raising by a small margin, and contrast technique 2 and 3 is it will be seen that the polysaccharide after depigmentation is again after uf processing, and polysaccharide content improves more, reaches 81%.
Table 4 depigmentation is on the impact (n=3) of 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 the determination of purifying process
According to table 3,4 result, it is determined 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 Integrated comparative 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 process, 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 processes, and this method is simple to operate, considers the pigment removal thought and be applicable to an authentic gas soup Crude polysaccharides. In addition, table 4 result shows: although depigmentation is less on the content impact of polysaccharide, but first depigmentation, then through ultrafiltration, polysaccharide content finally can reach more than 80%, depigmentation is step extremely important in an authentic gas soup polysaccharide purification technique as can be seen here.
As shown in Table 1, after an authentic gas soup is carried out deproteinated process, while albumen effective elimination, the content of polysaccharide also declines to some extent. But table 3 data show, after deproteinated process, polysaccharide content decline is many, all rises higher by the polysaccharide content after uf processing. Result illustrates, though the deproteinated process not polysaccharide content in raising Crude polysaccharides itself, this step is essential for the treating process of whole polysaccharide.
As shown in Table 4, Crude polysaccharides A, deproteinated Crude polysaccharides B are through ultrafiltration process, and after ultrafiltration process is refining, the content of polysaccharide is improved, and are easy to industrialization operation.
An authentic gas soup has more significant clinical efficacy, it is carried out deep Study on mechanism, illustrates its scientific meaning, have important clinical meaning. Particularly for the basic substance of its effect, it is 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. the method for the industrial extraction and purification of an authentic gas soup polysaccharide, it is characterised in that, comprise the steps: successively
1) taking 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 Radix Rehmanniae Preparata, all the other medicinal material dryings, pulverize;
2) by Radix Rehmanniae Preparata segment, with step 1) pulverize after medicinal material mix, by ultrasonic extraction extraction medicinal material 1-3 time, coarse filtration after each 7-9 times of water extraction 25-35min, 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, obtain Crude polysaccharides A in 55-65 DEG C of vacuum-drying after getting lower sediment washing with alcohol;
4) step 3 is got) Crude polysaccharides A is dissolved in water, and shakes even, and acid for adjusting pH, to 5-6, adds papoid, at 45-55 DEG C, enzymolysis 4-6h, deactivation, centrifugal removing precipitates, and gets subnatant and obtains Crude polysaccharides enzymolysis solution B;
5) to step 4) Crude polysaccharides enzymolysis solution B adds the chloroform-butanol solution of its volume 1/4-1/6, concussion, hold over night, collects upper strata aqueous phase solution, obtains polysaccharide soln B;
6) step 5 is got) polysaccharide soln B, add water and it is mixed with the solution that concentration is 3-5mg/ml, regulate the pH to 4.5-5.5 of polysaccharide soln, add macroporous resin and stir 1-3h under 45-55 DEG C of water-bath, filter to obtain Crude polysaccharides B;
7) step 6 is got) Crude polysaccharides B adds water and makes the solution that concentration is 0.15-0.25mg/mL, carries out ultrafiltration, collects and retains liquid, 55-65 DEG C of vacuum-drying after concentrated, obtains essence polysaccharide.
2. according to the method for industrial extraction and purification of an authentic gas soup polysaccharide according to claim 1, it is characterised in that, step 1) described in drying temperature be 60 DEG C.
3. the method for the industrial extraction and purification of an authentic gas soup polysaccharide according to claim 1, it is characterised in that, step 2) described in ultrasound condition stop 2s, power 800W at room temperature super 2s.
4. the method for the industrial extraction and purification of an authentic gas soup polysaccharide according to claim 1, it is characterised in that, step 2), step 3) and step 4) described in centrifugal condition be 5000rpm �� 10min.
5. the method for the industrial extraction and purification of an authentic gas soup polysaccharide according to claim 1, it is characterised in that, step 3) described in supernatant concentration 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, it is characterised in that, step 4) described in acid be Glacial acetic acid.
7. the method for the industrial extraction and purification of an authentic gas soup polysaccharide according to claim 1, it is characterised in that, get step 3) 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, it is characterised in that, step 4) described in deactivation be 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 in ultrafiltration charging flow velocity be 10-14mL/min, pressure is 0.08-012MPa so that be 1:1 through amount with the volume ratio of interception, when solution all enters ultrafiltration post, it is designated as ultrafiltration once, will retain in liquid and add 200mL distilled water, continue ultrafiltration, circulate after 25 times, collect and retain liquid.
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| 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 |
| 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 |
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| 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 |
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| KR101074158B1 (en) * | 2008-04-07 | 2011-10-17 | 재단법인 한국원자력의학원 | Composition comprising polysaccharide extracted from panax ginseng preventing and treating liver diseases |
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| 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 |
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