CN104045725B - D301G resin anion(R.A) is adopted to refine the method for Phaeopoms obliquus Crude polysaccharides - Google Patents
D301G resin anion(R.A) is adopted to refine the method for Phaeopoms obliquus Crude polysaccharides Download PDFInfo
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Abstract
The invention discloses a kind of method that the D301G of employing resin anion(R.A) refines Phaeopoms obliquus Crude polysaccharides, it is characterized in that, comprise the steps: the degreasing pre-treatment of A. raw material; B. hot water extraction prepares Crude polysaccharides; C. Crude polysaccharides enzymolysis; D. macroporous adsorbent resin D301G process; E. small molecules is removed in ultrafiltration; F. vacuum lyophilization; Prepare Fuscoporia obliqua polysaccharide.That use this technique finally to obtain decolouring and remove the Phaeopoms obliquus refined polysaccharide of most of albumen and small molecular weight impurity, polysaccharide retention rate is more than 58.2%, and percent of decolourization is 87.7%, and albumen decreasing ratio is up to 73.7%.Present method is applicable to industrialized scaleization and produces Phaeopoms obliquus refined polysaccharide.
Description
Technical field
The invention belongs to biological technical field, specifically relate to a kind of method that the D301G of employing resin anion(R.A) refines Phaeopoms obliquus Crude polysaccharides.
Background technology
Phaeopoms obliquus [
inonotusobliquus(Fr.) Pilat] be a famous Wild Medicinal fungi among the people.Research shows, Fuscoporia obliqua polysaccharide is one of important activeconstituents, has and strengthens immunomodulatory, the multiple pharmacologically active such as antitumor, anti-oxidant, hypoglycemic and antiviral, become one of hot fields of research both at home and abroad.But it is relatively less to the research report of the separation and purification of Fuscoporia obliqua polysaccharide at present.
Due to the composition complicacy of polysaccharide, the molecular weight distribution of polysaccharide crude extract is wider, polysaccharide kind is also a lot, and previous experiments shows, containing a certain amount of albumen in Phaeopoms obliquus Crude polysaccharides after extraction, pigment and small-molecule substance, affect the bioactive performance of polysaccharide and the further research of chemical structure, therefore be necessary the further separation and purification of Crude polysaccharides, but the impurity such as the albumen in removal Crude polysaccharides and pigment, it is a great problem in separation of polysaccharides purifying always, traditional Deproteinization is based on Sevage method, method complex operation, efficiency is low, the toxic biological activity thus affecting polysaccharide of organic solvent adopted, it is better that activated carbon method removes pigment effect, but the active carbon powder in later stage is not easily removed, and it is fine that hydrogen peroxide method removes pigment effect because of stronger anti-oxidant activity, but comparatively large to the activity influence of polysaccharide, is therefore badly in need of a kind of new process for purification of exploration.
Macroporous resin has macropore stereoscopic three-dimensional reticulated structure and larger specific surface area, therefore there is good exchange adsorption active, be widely used in the refining of the activeconstituents of natural product in recent years, some macroporous resin in order to remove albumen in plant and fungus polysaccharide and pigment, and achieves good effect.The research adopting macroporous resin to remove pigment and albumen in Phaeopoms obliquus Crude polysaccharides there is not yet bibliographical information.
Application publication number CN102603906A(application number 201110461635.8) Chinese patent literature disclose a kind of preparation method of the Fuscoporia obliqua polysaccharide aqueous solution: a kind of preparation method of the Fuscoporia obliqua polysaccharide aqueous solution, comprise the following steps: trichoroacetic acid(TCA) deproteinated: in Phaeopoms obliquus extract, add trichoroacetic acid(TCA), leave standstill, then solid-liquid separation, gets the solid matter being separated and obtaining; Activated carbon decolorizing: to step: gained solid matter by volume adds the water of 50 ~ 100 times, then adds activated carbon decolorizing, then solid-liquid separation, gets gained supernatant liquor; Clarify with natural clarifying agent: gained supernatant liquor adds natural clarifying agent clarification.
Summary of the invention
Order of the present invention is to provide a kind of technique of refining Phaeopoms obliquus Crude polysaccharides; core technology of the present invention is: applying biological enzyme and anion exchange absorbing resin substitute traditional Sevage method, activated carbon method and hydrogen peroxide method and remove albumen and pigment; overcome the defect of prior art, provide a kind of method that Phaeopoms obliquus refined polysaccharide is prepared in novel applicable mass-producing.
Technical scheme of the present invention is as follows:
Adopt D301G resin anion(R.A) to refine the method for Phaeopoms obliquus Crude polysaccharides, it comprises the steps: the degreasing pre-treatment of A. raw material; B. hot water extraction prepares Crude polysaccharides; C. Crude polysaccharides enzymolysis; D. macroporous resin D301G process; E. small molecules is removed in ultrafiltration; F. vacuum lyophilization; Prepare Phaeopoms obliquus refined polysaccharide.
Foregoing method, preferred scheme is, steps A. the degreasing pre-treatment of raw material refers to, gets dry inonotus obliquus sclerotium, pulverizes, it is preferred that the 60-95% ethanolic soln being placed in 4-8 times of volume soaks 16-24h(, the 70-90% ethanolic soln being placed in 6-7 times of volume soaks 18-20h, is more preferably, and 80% ethanolic soln being placed in 6.5 times of volumes soaks 20h), to remove degrease, part single (widow) sugar and fat-soluble pigment, residue cool place is located air-dry.
Foregoing method, preferred scheme is, step B. hot water extraction prepares Crude polysaccharides and refers to, after pretreated residue is air-dry, adds 10-20 times of distilled water (v/w), boiling water extraction 1-3h, 80-120 order filtered through gauze (preferred, add 13-18 times of distilled water (v/w), boiling water extraction 1.5-2.5h, 85-100 order filtered through gauze, be more preferably, add 15 times of distilled water (v/w), boiling water extraction 2h, 100 order filtered through gauze), filter residue repeats to extract 1-3 time, and merging filtrate is concentrated, centrifugal, spraying dry obtains Crude polysaccharides.
Foregoing method, preferred scheme is, step C. Crude polysaccharides enzymolysis refers to, is dissolved in water by Crude polysaccharides, add papoid, 35-45 DEG C of reaction 20-40min(is preferred, 36-42 DEG C of reaction 25-35min, be more preferably, 40 DEG C of reaction 30min), obtain the enzymolysis solution of Crude polysaccharides.Be more preferably, after Crude polysaccharides is dissolved in water, be formulated as 0.5-2.0%(w/w) the aqueous solution of the aqueous solution (preferred, be formulated as 0.6-1.8%(w/w), be more preferably, be formulated as 1.5%(w/w) the aqueous solution).Be more preferably, papoid add-on is the 3-5%(w/w of the Crude polysaccharides aqueous solution) (preferred, add-on is the 3.5-4.5%(w/w of the Crude polysaccharides aqueous solution), be more preferably, add-on is the 4.0%(w/w of the Crude polysaccharides aqueous solution)).
Foregoing method, preferred scheme is, step D. macroporous adsorbent resin D301G process refers to, the enzymolysis solution of Crude polysaccharides adds in pretreated D301G resin chromatography post, according to D301G Shu Zhi ︰ Crude polysaccharides enzymolysis solution=1.2-2.0 ︰ 1 (w/v), flow rate control is preferred at 1-3ml/min(, according to D301G Shu Zhi ︰ Crude polysaccharides enzymolysis solution=1.5-1.8 ︰ 1 (w/v), flow rate control is at 1.5-2ml/min, be more preferably, according to D301G Shu Zhi ︰ Crude polysaccharides enzymolysis solution=1.6 ︰ 1 (w/v), flow rate control is at 1.8ml/min), collect effluent liquid.Be more preferably, described pre-treatment refers to, macroporous adsorbent resin D301G is first soaked in water 24h, washes with water to the clarification of water liquid, inclines after anhydrating and add 1MHC1 solution soaking 24h, be washed to neutrality, add 1MNaOH solution soaking 24h, wash with water to neutrality.
Foregoing method, preferred scheme is, step e. ultrafiltration is removed small molecules and is referred to, adopt ultra-filtration membrane molecular weight be 10,000 hollow cellulose film carry out ultrafiltration, remove the small-molecule substance of less than 10,000.
Foregoing method, preferred scheme is, step F. vacuum lyophilization refers to, 3h at first Fuscoporia obliqua polysaccharide sample being positioned over-20 DEG C, then freezing more than 6h at proceeding to-80 DEG C, then proceeds to vacuum freeze drier drying, obtains refining Fuscoporia obliqua polysaccharide.
The invention discloses a kind of Phaeopoms obliquus refined polysaccharide and preparation technology thereof, preparation technology is as follows: 1, the pre-treatment of raw material; 2, the stripping of Phaeopoms obliquus Crude polysaccharides; 3, protease hydrolysis; 4, resin anion(R.A) process; 5, ultrafiltration, then dry.That use this technique finally to obtain decolouring and remove the Phaeopoms obliquus refined polysaccharide of most of albumen and small molecular weight impurity, polysaccharide retention rate is more than 58.2%, and percent of decolourization is 87.7%, and albumen decreasing ratio is up to 73.7%.Present method is applicable to industrialized scaleization and produces Phaeopoms obliquus refined polysaccharide.
Except this outside, technical superiority of the present invention is also embodied in: 1, adopt Papain ferment treatment Crude polysaccharides, improves the clearance (clearance up to 73.7%) of protein impurities, improves the purity of polysaccharide; 2, the refined polysaccharide adopting macroporous resin treatment to prepare, have no side effect, operate easier, purification effect is good, polysaccharide retention rate and yield high (respectively up to 58.2% and 62.4%).And Sevage method, chloroform used is toxic, and reclaims inconvenience, and yield is only 48.9%; Active carbon adsorption and hydrogen peroxide method remove pigment, all there is certain defect, active carbon adsorption bleaching time is grown and polysaccharide loss rate comparatively large (yield is only 48.6%), and gac is difficult to removing, hydrogen peroxide decolours, to the chemical structure of polysaccharide and biological activity greatly destructive.
Embodiment
Below in conjunction with embodiment, technical solution of the present invention is encyclopaedized, but protection domain is not by this restriction.Material used in embodiment is Phaeopoms obliquus Wild sclerotium, purchased from Heilungkiang institute of microbiology; The macroporous resin adopted is weakly basic styrene type anion exchange resin, and model is D301G, and purchased from recovery fine chemistry industry institute, particle size diameter is 0.60-1.60mm, creamy white; The present invention's proteolytic enzyme used is papoid, purchased from Beijing Ding Guo Bioisystech Co., Ltd.
embodiment 1adopt D301G resin anion(R.A) to refine the method for Phaeopoms obliquus Crude polysaccharides, comprise the steps:
A. the degreasing pre-treatment of raw material: get dry inonotus obliquus sclerotium, pulverize, 60% ethanolic soln being placed in 4 times of volumes soaks 16h, and to remove degrease, part single (widow) sugar and fat-soluble pigment, residue cool place is located air-dry.
B. hot water extraction prepares Crude polysaccharides: after pretreated residue is air-dry, adds 10 times of distilled water (v/w), boiling water extraction 1h, 80 order filtered through gauze, and filtrate concentrates, and centrifugal, spraying dry obtains Crude polysaccharides.
C. Crude polysaccharides enzymolysis: Crude polysaccharides is dissolved in water, is formulated as 0.5%(w/w) the aqueous solution, add papoid, papoid add-on is the 3%(w/w of the Crude polysaccharides aqueous solution), 35 DEG C reaction 20min, obtain the enzymolysis solution of Crude polysaccharides.
D. macroporous resin D301G process: the enzymolysis solution of Crude polysaccharides adds through pre-treatment that (macroporous adsorbent resin D301G is first soaked in water 24h, wash with water to water liquid and clarify, incline after anhydrating and add 1MHC1 solution soaking 24h, be washed to neutrality, add 1MNaOH solution soaking 24h, wash with water to neutrality) D301G resin chromatography post in, according to D301G Shu Zhi ︰ Crude polysaccharides enzymolysis solution=1.2 ︰ 1(w/v), flow rate control, at 1ml/min, collects effluent liquid.
E. small molecules is removed in ultrafiltration: adopt ultra-filtration membrane molecular weight be 10,000 hollow cellulose film carry out ultrafiltration, remove the small-molecule substance of less than 10,000, obtain Fuscoporia obliqua polysaccharide sample.
F. vacuum lyophilization: 3h at first Fuscoporia obliqua polysaccharide sample being positioned over-20 DEG C, freezing more than 6h at then proceeding to-80 DEG C, then proceeds to vacuum freeze drier drying, obtains refining Fuscoporia obliqua polysaccharide.
embodiment 2adopt D301G resin anion(R.A) to refine the method for Phaeopoms obliquus Crude polysaccharides, comprise the steps:
A. the degreasing pre-treatment of raw material: get dry inonotus obliquus sclerotium, pulverize, 95% ethanolic soln being placed in 8 times of volumes soaks 24h, and to remove degrease, part single (widow) sugar and fat-soluble pigment, residue cool place is located air-dry.
B. hot water extraction prepares Crude polysaccharides: after pretreated residue is air-dry, and add 20 times of distilled water (v/w), boiling water extraction 3h, 120 order filtered through gauze, filter residue repeats extraction 2 times, merging filtrate, and concentrated, centrifugal, spraying dry obtains Crude polysaccharides.
C. Crude polysaccharides enzymolysis: Crude polysaccharides is dissolved in water, is formulated as 2.0%(w/w) the aqueous solution, add papoid, papoid add-on is the 5%(w/w of the Crude polysaccharides aqueous solution), 45 DEG C reaction 40min, obtain the enzymolysis solution of Crude polysaccharides.
D. macroporous resin D301G process:
Macroporous adsorbent resin D301G pre-treatment, be first soaked in water 24h, washes with water to the clarification of water liquid, incline after anhydrating and add 1MHC1 solution soaking 24h, be washed to neutrality, add 1MNaOH solution soaking 24h, wash with water to neutrality.
The enzymolysis solution of Crude polysaccharides adds in pretreated D301G resin chromatography post, and according to D301G Shu Zhi ︰ Crude polysaccharides enzymolysis solution=2.0 ︰ 1 (w/v), flow rate control, at 3ml/min, collects effluent liquid.
E. small molecules is removed in ultrafiltration: adopt ultra-filtration membrane molecular weight be 10,000 hollow cellulose film carry out ultrafiltration, remove the small-molecule substance of less than 10,000.
F. vacuum lyophilization: 3h at first Fuscoporia obliqua polysaccharide sample being positioned over-20 DEG C, freezing more than 6h at then proceeding to-80 DEG C, then proceeds to vacuum freeze drier drying, obtains refining Fuscoporia obliqua polysaccharide.
embodiment 3adopt D301G resin anion(R.A) to refine the method for Phaeopoms obliquus Crude polysaccharides, it comprises the steps:
A. the degreasing pre-treatment of raw material: get dry inonotus obliquus sclerotium, pulverize, 90% ethanolic soln being placed in 7 times of volumes soaks 20h, and to remove degrease, part single (widow) sugar and fat-soluble pigment, residue cool place is located air-dry.
B. hot water extraction prepares Crude polysaccharides: after pretreated residue is air-dry, and add 18 times of distilled water (v/w), boiling water extraction 2.5h, 100 order filtered through gauze, filter residue repeats extraction 3 times, merging filtrate, and concentrated, centrifugal, spraying dry obtains Crude polysaccharides.
C. Crude polysaccharides enzymolysis: Crude polysaccharides is dissolved in water, is formulated as 1.8%(w/w) the aqueous solution, add papoid, papoid add-on is the 4.5%(w/w of the Crude polysaccharides aqueous solution), 42 DEG C reaction 35min, obtain the enzymolysis solution of Crude polysaccharides.
D. macroporous resin D301G process: the enzymolysis solution of Crude polysaccharides adds in the D301G resin chromatography post of pre-treatment (pretreatment mode be that embodiment 1-3 is identical), according to D301G Shu Zhi ︰ Crude polysaccharides enzymolysis solution=1.8 ︰ 1 (w/v), flow rate control, at 2ml/min, collects effluent liquid.
E. small molecules is removed in ultrafiltration: adopt ultra-filtration membrane molecular weight be 10,000 hollow cellulose film carry out ultrafiltration, remove the small-molecule substance of less than 10,000.
F. vacuum lyophilization: 3h at first Fuscoporia obliqua polysaccharide sample being positioned over-20 DEG C, freezing more than 6h at then proceeding to-80 DEG C, then proceeds to vacuum freeze drier drying, obtains refining Fuscoporia obliqua polysaccharide.
embodiment 4adopt D301G resin anion(R.A) to refine the method for Phaeopoms obliquus Crude polysaccharides, comprise the steps:
A. the degreasing pre-treatment of raw material: get dry inonotus obliquus sclerotium, pulverize, 70% ethanolic soln being placed in 6 times of volumes soaks 18h, and to remove degrease, part single (widow) sugar and fat-soluble pigment, residue cool place is located air-dry.
B. hot water extraction prepares Crude polysaccharides: after pretreated residue is air-dry, adds 13 times of distilled water (v/w), boiling water extraction 1.5h, 85 order filtered through gauze, extracts 1 time, and filtrate concentrates, and centrifugal, spraying dry obtains Crude polysaccharides.
C. Crude polysaccharides enzymolysis: Crude polysaccharides is dissolved in water, is formulated as 0.5%(w/w) the aqueous solution, add papoid, papoid add-on is the 3.5%(w/w of the Crude polysaccharides aqueous solution), 36 DEG C reaction 25min, obtain the enzymolysis solution of Crude polysaccharides.
D. macroporous resin D301G process: the enzymolysis solution of Crude polysaccharides adds through pre-treatment that (macroporous adsorbent resin D301G is first soaked in water 24h, wash with water to water liquid and clarify, incline after anhydrating and add 1MHC1 solution soaking 24h, be washed to neutrality, add 1MNaOH solution soaking 24h, wash with water to neutrality) D301G resin chromatography post in, according to D301G Shu Zhi ︰ Crude polysaccharides enzymolysis solution=1.5 ︰ 1(w/v), flow rate control, at 1.5ml/min, collects effluent liquid.
E. small molecules is removed in ultrafiltration: adopt ultra-filtration membrane molecular weight be 10,000 hollow cellulose film carry out ultrafiltration, remove the small-molecule substance of less than 10,000.
F. vacuum lyophilization: 3h at first Fuscoporia obliqua polysaccharide sample being positioned over-20 DEG C, freezing more than 6h at then proceeding to-80 DEG C, then proceeds to vacuum freeze drier drying, obtains refining Fuscoporia obliqua polysaccharide.
embodiment 5adopt D301G resin anion(R.A) to refine the method for Phaeopoms obliquus Crude polysaccharides, it comprises the steps:
A. the degreasing pre-treatment of raw material: get dry inonotus obliquus sclerotium, pulverize, 80% ethanol (mass concentration) solution being placed in 6.5 times of volumes soaks 20h, and to remove degrease, part oligosaccharides and fat-soluble pigment, residue cool place place is air-dry.
B. hot water extraction prepares Crude polysaccharides: after pretreated residue is air-dry, and add 15 times of distilled water (v/w), boiling water extraction 2h, 100 order filtered through gauze, filter residue repeats extraction 2 times, merging filtrate, and concentrated, centrifugal, spraying dry obtains Crude polysaccharides.
C. Crude polysaccharides enzymolysis: Crude polysaccharides is dissolved in water, is formulated as 1.5%(w/w) the aqueous solution, add papoid, papoid add-on is 4.0% (w/w) of the Crude polysaccharides aqueous solution, 40 DEG C reaction 30min, obtain the enzymolysis solution of Crude polysaccharides.
D. macroporous resin D301G process: the enzymolysis solution of Crude polysaccharides adds through pre-treatment that (described pre-treatment refers to, macroporous adsorbent resin D301G is first soaked in water 24h, wash with water to water liquid and clarify, incline after anhydrating and add 1MHC1 solution soaking 24h, be washed to neutrality, add 1MNaOH solution soaking 24h, wash with water to neutrality) D301G resin chromatography post in, according to D301G Shu Zhi ︰ Crude polysaccharides enzymolysis solution=1.6 ︰ 1 (w/v), flow rate control, at 1.8ml/min, collects effluent liquid.
E. small molecules is removed in ultrafiltration: adopt ultra-filtration membrane molecular weight be 10,000 hollow cellulose film carry out ultrafiltration, remove the small-molecule substance of less than 10,000.
F. vacuum lyophilization: 3h at first Fuscoporia obliqua polysaccharide sample being positioned over-20 DEG C, freezing more than 6h at then proceeding to-80 DEG C, then proceeds to vacuum freeze drier drying, obtains refining Fuscoporia obliqua polysaccharide.
test examplethe mensuration of embodiment 5 gained Fuscoporia obliqua polysaccharide polysaccharide content and protein content, and the calculating of albumen decreasing ratio, pigment removal rate and polysaccharide retention rate.The wherein content (%) of content (the %)-reducing sugar of polysaccharide content (%)=total reducing sugar.
Total sugar content measures and adopts phend-sulphuric acid; Reducing sugar content measures and adopts DNS method; Determining the protein quantity adopts Bradford method.
In formula: A
0, A
1solution absorbancy at wavelength 450nm place rear with decolouring before being respectively decolouring.
In formula: C
0, C
1solution absorbancy at wavelength 450nm place rear with decolouring before being respectively decolouring.
In formula: M
0, M
1be respectively the polysaccharide content before and after process.
Here is the macroporous resin D301G(embodiment of the present invention 5) refine the results contrast of Phaeopoms obliquus Crude polysaccharides with Sevage method, activated carbon method and hydrogen peroxide method, this shows, macroporous resin D301G, have the ability removing albumen and pigment concurrently.See the following form:
The present invention adopts D301G resin anion(R.A) to refine the method for Phaeopoms obliquus Crude polysaccharides, not only albumen can be removed, also have the ability removing pigment concurrently, therefore the purity of polysaccharide prepared is higher, compared with the Crude polysaccharides before process, pigment removal rate is 87.7%, albumen decreasing ratio is 73.7%, polysaccharide retention rate 58.2%.
The application is subsidized by Shandong Province's Nsfc Projects (ZR2010CL008).
Finally it should be noted that, embodiment is the embodiment of optimum of the present invention, be not limited to the present invention, although with reference to previous embodiment to invention has been detailed description, for a person skilled in the art, it still can be modified to the technical scheme described in foregoing embodiments, or carries out equivalent replacement to wherein portion of techniques feature.Within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (13)
1. adopt D301G resin anion(R.A) to refine the method for Phaeopoms obliquus Crude polysaccharides, it is characterized in that, comprise the steps:
A. the degreasing pre-treatment of raw material: get dry inonotus obliquus sclerotium, pulverize, the 60-95% ethanolic soln being placed in 4-8 times of volume soaks 16-24h, and to remove degrease, part monose and fat-soluble pigment, residue cool place place is air-dry;
B. hot water extraction prepares Crude polysaccharides: after pretreated residue is air-dry, adds 10-20 times of distilled water (v/w), boiling water extraction 1-3h, 80-120 order filtered through gauze, and filter residue repeats to extract 1-3 time, merging filtrate, and concentrated, centrifugal, spraying dry obtains Crude polysaccharides;
C. Crude polysaccharides enzymolysis: be formulated as 0.5-2.0%(w/w after Crude polysaccharides is dissolved in water) the aqueous solution, add papoid, papoid add-on is the 3-5%(w/w of the Crude polysaccharides aqueous solution), 35-45 DEG C reaction 20-40min, obtain the enzymolysis solution of Crude polysaccharides;
D. macroporous adsorbent resin D301G process: the enzymolysis solution of Crude polysaccharides adds in pretreated D301G resin chromatography post, according to D301G Shu Zhi ︰ Crude polysaccharides enzymolysis solution=1.2-2.0 ︰ 1(w/v), flow rate control, at 1-3ml/min, collects effluent liquid; Described pre-treatment refers to, macroporous adsorbent resin D301G is first soaked in water 24h, washes with water to the clarification of water liquid, inclines after anhydrating and add 1MHC1 solution soaking 24h, be washed to neutrality, add 1MNaOH solution soaking 24h, wash with water to neutrality;
E. small molecules is removed in ultrafiltration: adopt ultra-filtration membrane molecular weight be 10,000 hollow cellulose film carry out ultrafiltration, remove the small-molecule substance of less than 10,000, obtain Fuscoporia obliqua polysaccharide sample;
F. vacuum lyophilization; Prepare Fuscoporia obliqua polysaccharide: described vacuum lyophilization refers to, 3h at first Fuscoporia obliqua polysaccharide sample being positioned over-20 DEG C, freezing more than 6h at then proceeding to-80 DEG C, then proceeds to vacuum freeze drier drying, obtains refining Fuscoporia obliqua polysaccharide.
2. method according to claim 1, is characterized in that, steps A: the 70-90% ethanolic soln being placed in 6-7 times of volume soaks 18-20h, and to remove degrease, part monose and fat-soluble pigment, residue cool place place is air-dry.
3. method according to claim 2, is characterized in that, steps A: 80% ethanolic soln being placed in 6.5 times of volumes soaks 20h.
4. method according to claim 1, is characterized in that, step B: add 13-18 times of distilled water (v/w), boiling water extraction 1.5-2.5h, 85-100 order filtered through gauze, and filter residue repeats to extract 1-3 time, merging filtrate, and concentrated, centrifugal, spraying dry obtains Crude polysaccharides.
5. method according to claim 4, is characterized in that, step B: add 15 times of distilled water (v/w), boiling water extraction 2h, 100 order filtered through gauze.
6. method according to claim 1, is characterized in that, step C:36-42 DEG C of reaction 25-35min, obtains the enzymolysis solution of Crude polysaccharides.
7. method according to claim 6, is characterized in that, step C:40 DEG C of reaction 30min.
8. method according to claim 1, is characterized in that, step C: be formulated as 0.6-1.8%(w/w after Crude polysaccharides is dissolved in water) the aqueous solution.
9. method according to claim 8, is characterized in that, step C: be formulated as 1.5%(w/w after Crude polysaccharides is dissolved in water) the aqueous solution.
10. method according to claim 1, is characterized in that, step C: papoid add-on is the 3.5-4.5%(w/w of the Crude polysaccharides aqueous solution).
11. methods according to claim 10, is characterized in that, step C: papoid add-on is the 4.0%(w/w of the Crude polysaccharides aqueous solution).
12. methods according to claim 1, is characterized in that, step D: according to D301G Shu Zhi ︰ Crude polysaccharides enzymolysis solution=1.5-1.8 ︰ 1(w/v), flow rate control, at 1.5-2ml/min, collects effluent liquid.
13. methods according to claim 12, is characterized in that, step D: according to D301G Shu Zhi ︰ Crude polysaccharides enzymolysis solution=1.6 ︰ 1(w/v), flow rate control is at 1.8ml/min.
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| CN110150668B (en) * | 2019-05-30 | 2022-09-23 | 青岛农业大学 | Inonotus obliquus enzymolysis polysaccharide oral liquid |
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| CN102038720A (en) * | 2010-12-17 | 2011-05-04 | 中国农业大学 | Fuscoporia obliqua active ingredients capable of lowering blood sugar and preparation method and application of fuscoporia obliqua active ingredients |
| CN102603906A (en) * | 2011-12-30 | 2012-07-25 | 安徽丰原发酵技术工程研究有限公司 | Preparation method of inonotus obliquus polysaccharide aqueous solution |
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| KR100340663B1 (en) * | 2000-12-16 | 2002-06-20 | 정종상 | A method of preparing the antitumor and immuno-activity polysaccharide from the artificial cultivation of inonotus obliquus, and its use |
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2014
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102038720A (en) * | 2010-12-17 | 2011-05-04 | 中国农业大学 | Fuscoporia obliqua active ingredients capable of lowering blood sugar and preparation method and application of fuscoporia obliqua active ingredients |
| CN102603906A (en) * | 2011-12-30 | 2012-07-25 | 安徽丰原发酵技术工程研究有限公司 | Preparation method of inonotus obliquus polysaccharide aqueous solution |
Non-Patent Citations (1)
| Title |
|---|
| 两种不同来源桦褐孔菌多糖的抗氧化与免疫活性的研究;武永德;《中国优秀硕士学位论文全文数据库（医药卫生科技辑）》;20120315(第3期);第26-27页 * |
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