CN109134688A - A kind of galactomannans and preparation method thereof rich in furans configuration - Google Patents
A kind of galactomannans and preparation method thereof rich in furans configuration Download PDFInfo
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- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0087—Glucomannans or galactomannans; Tara or tara gum, i.e. D-mannose and D-galactose units, e.g. from Cesalpinia spinosa; Tamarind gum, i.e. D-galactose, D-glucose and D-xylose units, e.g. from Tamarindus indica; Gum Arabic, i.e. L-arabinose, L-rhamnose, D-galactose and D-glucuronic acid units, e.g. from Acacia Senegal or Acacia Seyal; Derivatives thereof
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Abstract
The present invention relates to the preparation methods of galactomannans, for the polysaccharide for spending the higher furans configuration of middle preparation activity from golden cicada, disclose the galactomannans and preparation method thereof rich in furans configuration, the connection type of the strand of the galactomannans are as follows: → 2)-α-D-Manp (1 →, → 6)-α-D-Manp (1 →, → 4)-α-D-Glcp (1 →, → 2, 4)-α-D-Glcp (1 →, → 5)-β-D-Galf (1 →, → 2)-β-D-Galf (1 →, , molar ratio is 1:1.3:8:2:3:2, the flooding after golden cicada flower sporinite degreasing, it is concentrated under reduced pressure, ethanol solution precipitating, removal of protein, it is freeze-dried after desalination, then through Q Sepharose TMFa Elution, evaporation and concentration obtain after st Flow post separation, and the accounting of furans configuration reaches 28.9% in galactomannans, and extraction efficiency is high.
Description
Technical field
The present invention relates to the preparation methods of galactomannans, and in particular to a kind of galactomannan rich in furans configuration is poly-
Sugar and preparation method thereof.
Background technique
Furan derivatives are the important compound and intermediate of field of medicaments, have antibacterial and other effects.And there is furans structure
The glycan molecule of type not only has the characteristics that chirality, hydrophily, lipophilicity and flexibility, also has absolute steric configuration, therefore
It can be used as chiral auxiliary and chiral building block, to synthesize the asymmetric molecult with very strong bioactivity.
Cicada fungus is a kind of assembly that entomogenous fungi is interdependent, similar cordyceps sinensis, and forming process is that the larva of cicada sprouts wings in cicada
It is preceding by Cordyceps Militaris infection, it is parasitic, when climatic environment is suitable for, the nutrition for absorbing polypide is converted to mycelium, and from the vegetative phase
" the cicada fungus conidia powder " that there is breeding function for the perfect stage is gradated, is similar to flower from top branch " germination ", so claim
For cicada fungus, including originate in " the golden cicada flower " in Zhejiang, " the small cicada fungus " of " cicada fungus " in Sichuan and Guangdong, Fujian " native cicada fungus ", with
And " the only horned dinosaur " or " black the top of the horn " of Ningbo of Zhejiang.Cicada fungus belongs to high-quality cordyceps sinensis, has obvious anti-aging, immunological regulation, resists and answer
Sharp, antifatigue, anti anoxia and other effects.Recent study is found in cicada fungus not only containing protein, fat and a small amount of micro member
Element, polysaccharide also rich in.How from golden cicada to spend middle polysaccharide of the preparation containing furans configuration, expands golden cicada flower and is preparing drug
Application in terms of compound and important intermediate is of great significance to the further value for promoting golden cicada flower.
Chinese invention patent application CN201410232441.4, denomination of invention golden cicada spend polysaccharide and its are preparing neuroprotection
With the application in antiaging agent, discloses with golden cicada flower as raw material, after water extracts back, dehydrated alcohol is added to precipitate, then
Centrifugal drying precipitates to obtain the preparation method of golden cicada flower polysaccharide, but the golden cicada flower polysaccharide of this method preparation is unable to ensure containing furan
It mutters configuration.
Summary of the invention
To spend the middle polysaccharide for preparing furans configuration from golden cicada, the purpose of the present invention is to provide a kind of rich in furans configuration
Galactomannans, the galactomannans are prepared from golden cicada flower, and rare furans configuration is contained in molecular structure.
Another object of the present invention is to provide a kind of preparation methods of galactomannans that should be rich in furans configuration.
The present invention provides the following technical solution:
A kind of galactomannans rich in furans configuration, galactomannans isolate and purify to obtain from golden cicada flower sporinite,
It is made of mannose, glucose sugar, galactolipin and arabinose 1:5:2.5:1 polymerization in molar ratio, molecular weight 1.8kDa.
As a preference of the present invention, the strand connection type of the galactomannans are as follows: → 2)-α-D-Manp (1
→,→6)-α-D-Manp(1→,→4)-α-D-Glcp(1→,→2,4)-α-D-Glcp(1→,→5)-β-D-Galf(1→,
→ 2)-β-D-Galf (1 →, molar ratio is followed successively by 1:1.3:8:2:3:2.
Galactomannans of the invention is separated from golden cicada flower sporinite and is prepared, and has furans configuration, i.e. β-D-
Galf, the molar ratio in strand reach 28.9%, rich content.Simultaneously the galactomannans by mannose, glucose,
Galactolipin and arabinose composition, molecular weight is low, is easy to apply in medicine synthesis, there is wider application prospect.
The preparation method of the above-mentioned galactomannans rich in furans configuration, comprising the following steps:
(1) by golden cicada flower sporinite grinding and sieving, acetone degreasing；
(2) the golden cicada flower sporinite after extracting degreasing extracts after adding water, is then centrifuged for obtaining supernatant；
(3) sporinite is spent as step (2) extract golden cicada repeatedly, and the supernatant merged after centrifugation obtains extracting solution；
(4) extracting solution obtained by step (3) is concentrated under reduced pressure, low-concentration ethanol solution precipitating is first added, after being centrifuged removal precipitating
High concentration ethanol solution precipitating is added, is centrifugally separating to obtain head product；
(5) head product after separation is dissolved with water, successively deproteination matter, dialysis desalination, are then freeze-dried to obtain raw sugar；
(6) raw sugar dissolution is placed on strong anion exchange column Q Sepharose TMFast Flow and is separated；
(7) using sodium chloride solution, successively it is maximum to choose absorbance for gradient elution, the simultaneously sulfuric acid-phynol method detection of collection eluent
Eluting peak where eluent be concentrated by evaporation, be dried to obtain final product.
The galactomannans rich in furans configuration flooding after golden cicada flower sporinite degreasing, after being then concentrated under reduced pressure
Ethanol solution precipitating, then freeze-drying obtains raw sugar after deproteination matter, desalination, then through strong anion exchange column Q
It is eluted after Sepharose TMFast Flow separation, and is concentrated by evaporation eluent and obtains final product.During elution, to sulphur
Product in the corresponding eluent of the detected difference eluting peak of acid-phynol method is further detected, true by analyzing
Determining the polysaccharide product in the maximum eluent of eluting peak is the galactomannans rich in furans configuration, opposite golden cicada flower sporinite
Yield is 10% or so.
As the preferred of the method for the present invention, in leaching process, the mass volume ratio 1g:20 of golden cicada flower sporinite and water~
30mL, is extracted 2~5 times repeatedly by 96~100 DEG C of extraction temperature, 4~5h of extraction time.Extraction it is slightly boiled under fluidized state into
On the one hand row accelerates leaching rate, on the other hand carry the part volatile substances in golden cicada flower sporinite secretly ease by boiling
Out, the denaturation for promoting protein improves the purity of final product, and more preferable by the bactericidal effect of boiling extraction.
As the preferred of the method for the present invention, the concentration of low-concentration ethanol solution is 40wt%~45wt% in step (4), is added
Volume is 4~6 times that leaching liquor is concentrated, and high concentration ethanol solution concentration is 77wt%~80wt%, and it is removal precipitating that volume, which is added,
4~6 times of solution afterwards.Inventor has found in an experiment, selects the ethyl alcohol of higher concentration or the resulting impurity level of straight alcohol
Greatly, type is more, and uses the ethanol solution of low concentration then relatively more to the extraction of other polysaccharide, especially compared with final product
The big polysaccharide of molecular weight.Therefore it explores through a large number of experiments, just the preparation of the galactomannans rich in furans configuration is directed to
Property first using the polysaccharide of the ethanol solution sedimentation macromolecule of 40wt%~45wt%, then using concentration 77wt%~80wt%
Ethanol solution precipitates galactomannans, and precipitating will not settle.
As the preferred of the method for the present invention, Sevag method deproteination matter is used in step (5), using molecular cut off
The bag filter dialysis desalination of 3500Da.
As the preferred of the method for the present invention, the separation process of step (6) is as follows: the raw sugar product of 50mg is dissolved in 1~
In the deionized water of 2mL, then 0.45 μm of micro-filtrate membrane filtration, then loading separation, loading speed is 1mL/min.The study found that mistake
The purification efficiency of low loading speed is low, excessively high, causes not adsorb.
As the preferred of the method for the present invention, the elution process of step (7) is as follows: various concentration NaCl solution is pressed from low dense
It spends and successively divides gradient elution splitter to high concentration, NaCl solution concentration gradient are as follows: 0,0.1 mol/L, 0.25 mol/L, 0.5
Mol/L, elution flow rate 3mL/min, each gradient collect the eluent of 2~3 times of column volumes.Flow velocity is to elution effect shadow at this time
Sound is smaller.
As the preferred of the method for the present invention, the wavelength of sulfuric acid-phynol method detection is 490nm.
Beneficial effects of the present invention are as follows:
Galactomannans of the invention furans configuration rich in, the molar ratio of furans configuration reach 28.9%, and molecule
It measures low.The galactomannans separates preparation from golden cicada flower sporinite, and extraction efficiency is high, and the sweet glycan of gala is with respect to golden cicada flower spore
The yield of daughter is 10% or so, and yield is higher.
Detailed description of the invention
Fig. 1 is the molecular chain structure figure of galactomannans of the invention.
Fig. 2 is the light absorption value elution curve that sulfuric acid-phynol detection method of the invention obtains.
Fig. 3 is the canonical plotting mapped as ordinate to chromatographic retention using standard glucose molecular weight logarithm.
Fig. 4 is the high performance liquid chromatography retention time spectrogram of final product polysaccharide.
Fig. 5 is each monosaccharide high performance liquid chromatography separation spectrogram of the final product polysaccharide after PMP derivative.
Fig. 6 is the carbon-13 nmr spectra spectrogram of final product polysaccharide.
In Fig. 2, a indicates that the relation curve of absorbance and elution volume, b indicate NaCl solution concentration-gradient curve.
Specific embodiment
A specific embodiment of the invention is described further below.
Unless otherwise instructed, raw material employed in the present invention is commercially available or commonly used in the art, such as
Without special instruction, the method in following embodiments is the conventional method of this field.
Embodiment 1
A kind of galactomannans rich in furans configuration isolates and purifies to obtain, by mannose, glucose from golden cicada flower sporinite
Sugar, galactolipin and the arabinose composition of 1:5:2.5:1 polymerization in molar ratio, molecular weight 1.8kDa, strand connection type are as follows:
→2)-α-D-Manp(1→,→6)-α-D-Manp(1→,→4)-α-D-Glcp(1→,→2,4)-α-D-Glcp(1→,→
5)-β-D-Galf (1 →, → 2)-β-D-Galf (1 →, molar ratio shared by each configuration is 1:1.3:8:2:3:2, and the gala is sweet
The molecular chain structure figure for revealing glycan is as shown in Figure 1.
Embodiment 2
A kind of preparation method of the galactomannans rich in furans configuration in embodiment 1, comprising the following steps:
(1) 40 mesh sieves, acetone degreasing after crushing golden cicada flower sporinite, then 40 DEG C of dryings；
(2) the golden cicada flower sporinite 1g after extracting degreasing adds 20mL flooding 4h, 96 DEG C of extraction temperature, is then centrifuged for obtaining supernatant；
(3) it is spent sporinite 2 times as step (2) extract golden cicada repeatedly, and the supernatant merged after centrifugation obtains extracting solution；
(4) extracting solution obtained by step (3), first plus low-concentration ethanol solution precipitates overnight, low-concentration ethanol solution is concentrated under reduced pressure
Concentration is 45wt%, and it is 4 times that leaching liquor is concentrated that volume, which is added, and centrifugation removal precipitates, then high concentration ethanol solution precipitating, high
Concentration ethanol solution concentration is 77wt%, and 4 times that volume is the solution after removal precipitating are added, are centrifugally separating to obtain head product；
(5) head product after separation is dissolved with water, successively using Sevag method deproteination matter, using molecular cut off
The bag filter dialysis desalination of 3500Da, is then freeze-dried to obtain raw sugar；
(6) 50mg raw sugar is dissolved in the deionized water of 1mL, then 0.45 μm of micro-filtrate membrane filtration, then filtrate is placed in strong yin
It is separated on ion exchange column Q Sepharose TMFast Flow, loading speed is 1ml/min；
(7) using sodium chloride solution from low concentration to high concentration successively gradient elution, NaCl solution concentration gradient are as follows: 0,0.1
Mol/L, 0.25 mol/L, 0.5 mol/L, elution flow rate 3mL/min, each gradient collect the eluent of 2 times of column volumes, and
Sulfuric acid-phynol method detection, the wavelength of detection are 490nm, then choose the eluent evaporation where the maximum eluting peak of absorbance
It is concentrated, is dried to obtain final product polysaccharide, the recovery rate of opposite golden cicada flower steamed stuffed bun body is 10.2%.
Embodiment 3
A kind of preparation method of the galactomannans rich in furans configuration in embodiment 1, comprising the following steps:
(1) 40 mesh sieves, acetone degreasing after crushing golden cicada flower sporinite, then 40 DEG C of dryings；
(2) the golden cicada flower sporinite 1g after extracting degreasing adds 30mL flooding 5h, 100 DEG C of extraction temperature, is then centrifuged for obtaining supernatant；
(3) it is spent sporinite 3 times as step (2) extract golden cicada repeatedly, and the supernatant merged after centrifugation obtains extracting solution；
(4) extracting solution obtained by step (3) is concentrated under reduced pressure, low-concentration ethanol solution precipitates overnight, low-concentration ethanol is first added
Solution concentration is 45wt%, and it is 5 times that leaching liquor is concentrated that volume, which is added, and it is heavy that high concentration ethanol solution is added after centrifugation removal precipitating
It forms sediment, high concentration ethanol solution concentration is 80wt%, and 5 times that volume is the solution after removal precipitating are added, are centrifugally separating to obtain primiparity
Object；
(5) head product after separation is dissolved with water, successively using Sevag method deproteination matter, using molecular cut off
The bag filter dialysis desalination of 3500Da, is then freeze-dried to obtain raw sugar；
(6) 50mg raw sugar is dissolved in the deionized water of 2mL, then 0.45 μm of micro-filtrate membrane filtration, then filtrate is placed in strong yin
It is separated on ion exchange column Q Sepharose TMFast Flow, loading speed is 1ml/min；
(7) using sodium chloride solution from low concentration to high concentration successively gradient elution, NaCl solution concentration gradient are as follows: 0,0.1
Mol/L, 0.25 mol/L, 0.5 mol/L, elution flow rate 3mL/min, each gradient collect the eluent of 3 times of column volumes, and
Sulfuric acid-phynol method detection, the wavelength of detection are 490nm, then choose the eluent evaporation where the maximum eluting peak of absorbance
It is concentrated, is dried to obtain final product polysaccharide, the recovery rate of opposite golden cicada flower sporinite is 10.5%.
Embodiment 4
A kind of preparation method of the galactomannans rich in furans configuration in embodiment 1, comprising the following steps:
(1) 40 mesh sieves, acetone degreasing after crushing golden cicada flower sporinite, then 40 DEG C of dryings；
(2) the golden cicada flower sporinite 1g after extracting degreasing adds 25mL flooding 4h, 98 DEG C of extraction temperature, is then centrifuged for obtaining supernatant；
(3) it is spent sporinite 5 times as step (2) extract golden cicada repeatedly, and the supernatant merged after centrifugation obtains extracting solution；
(4) extracting solution obtained by step (3) is concentrated under reduced pressure, low-concentration ethanol solution precipitates overnight, low-concentration ethanol is first added
Solution concentration is 43wt%, and it is 6 times that leaching liquor is concentrated that volume, which is added, and it is heavy that high concentration ethanol solution is added after centrifugation removal precipitating
It forms sediment, high concentration ethanol solution concentration is 78wt%, and 6 times that volume is the solution after removal precipitating are added, are centrifugally separating to obtain primiparity
Object；
(5) head product after separation is dissolved with water, successively using Sevag method deproteination matter, using molecular cut off
The bag filter dialysis desalination of 3500Da, is then freeze-dried to obtain raw sugar；
(6) 50mg raw sugar is dissolved in the deionized water of 2mL, then 0.45 μm of micro-filtrate membrane filtration, then filtrate is placed in strong yin
It is separated on ion exchange column Q Sepharose TMFast Flow, loading speed is 1ml/min；
(7) using sodium chloride solution from low concentration to high concentration successively gradient elution, NaCl solution concentration gradient are as follows: 0,0.1
Mol/L, 0.25 mol/L, 0.5 mol/L, elution flow rate 3mL/min, each gradient collect the eluent of 2 times of column volumes, and
Sulfuric acid-phynol method detection, the wavelength of detection are 490nm, then choose the eluent evaporation where the maximum eluting peak of absorbance
It is concentrated, is dried to obtain final product polysaccharide, the recovery rate of opposite golden cicada flower sporinite is 10.6%.
Indexs measure is carried out by object of the final product polysaccharide of embodiment 2.
(1) the sulfuric acid-phynol method detection of raw sugar eluent
As shown in 2 step of embodiment (6) and step (7), the raw sugar product of 50mg is dissolved in the deionized water of 1mL, then
0.45 μm of micro-filtrate membrane filtration, then be placed on strong anion exchange column Q Sepharose TMFast Flow and separate, loading speed
1ml/min；Then be 0,0.1 mol/L by concentration gradient, the NaCl solution of 0.25 mol/L, 0.5 mol/L are pressed from low concentration
To high concentration successively gradient elution splitter, the eluent of 2 times of column volumes is collected, then using sulfuric acid-phynol method measurement elution
The light absorption value size of liquid, wavelength 490nm, according to light absorption value relative elutropic liquid volume rendering elution curve, as a result such as Fig. 2 institute
Show, a indicates that the relation curve of absorbance and elution volume, b indicate NaCl solution concentration-gradient curve in Fig. 2.It can be with from figure
Find out, pure water elutes the available maximum eluting peak of light absorption value.
(2) molecular weight determination
Using the molecular weight of High Performance Gel Permeation Chromatography measurement final product, chromatographic condition are as follows: 1260 liquid chromatogram of Agilent
Instrument；Analytical column: SB-HQ-804 chromatographic column；Mobile phase: 0.1 mol/L Na2SO4Aqueous solution；Flow velocity: 0.5 mL/min；Column temperature:
35 ℃；Sample volume: 20 μ L；Detector: Composition distribution RI Detector.
Each glucan serial standards are made into 5 mg/mL aqueous solutions with mobile phase, with pair of dextran standards molecular weight
Number (log Mw) is to chromatographic retention (tR) canonical plotting of mapping to obtain, as a result as shown in Figure 3.Then final product polysaccharide is used
0.1 mol/L Na2SO4The concentration that flowing phased soln is configured to 5 mg/mL carries out high-efficient liquid phase analysis, gained spectrogram such as Fig. 4 institute
Show, determines that retention time is 14.1min；The relative molecular weight of final product is calculated using standard curve, molecular weight is 1.8 kDa.
(3) monosaccharide composition analysis of final product polysaccharide
Using PMP column front derivation efficient liquid phase color method analysis monosaccharide composition after complete sour water solution.PMP derivatization method is used first
Product after the monosaccharide standard and final product polysaccharide hydrolysis prepare to equimolar carries out PMP and derives, and then carries out efficient liquid phase
Chromatography, as a result as shown in Figure 5.It can be seen from the figure that the monosaccharide composition and molar ratio of the galactomannans are sweet dew
Sugar: glucose: galactolipin: arabinose=1:5:2.5:1 is a kind of neutral sugar based on glucose and galactolipin.
(4) the configuration detection of final product polysaccharide
Carbon-13 nmr spectra analysis is carried out to obtained final product polysaccharide, as a result as shown in fig. 6,109.4ppm and 108.6 in figure
Ppm shows the signal of galactofuranose.
Methylation analysis is carried out to final product polysaccharide is obtained using the Hakomori method of improvement: weighing 1.0 mg and is dried under reduced pressure
1.0 mL dimethyl sulfoxides are added in final product polysaccharide afterwards, and magnetic agitation to sample is completely dissolved, and it is dry to rapidly join 100.0 mg
Dry NaH, is filled with N2Reacted at room temperature after sealing 1 hour, be slowly added to 1.0 mL of iodomethane, the reaction was continued 1 hour, finally
1.0 mL water are added and terminate reaction, reaction solution is multiple with chloroform extraction, combining extraction liquid, and three times with water washing chloroform layer.It will
Polysaccharide is dried under reduced pressure after methylation, does infrared spectrum analysis to determine whether methylation is complete, then by the end after methylation
The complete sour water solution of product polysaccharide sample, sodium borohydride reduction, then acetyl derivatives are obtained by acetic anhydride acetylation, then carry out
The strand connection type of efficient Gc-mss, gained final product polysaccharide is as shown in Figure 1.
Determined by above-mentioned analysis, the connection type of gained final product polysaccharide galactomannans is → 2)-α-D-Manp
(1→, →6)-α-D-Manp(1→, →4)-α-D-Glcp(1→, →2,4)-α-D-Glcp(1→, →5)-β-D-
Galf (1 →, → 2)-β-D-Galf (1 →, molar ratio 1:1.3:8:2:3:2, mole accounting of furans configuration β-D-Galf
It is 28.9%.
It should be noted that the final product of embodiment 3 and embodiment 4 is identical as the molecular weight of embodiment 2 and configuration, it is different
Place is recovery rate difference.
Claims (9)
1. a kind of galactomannans rich in furans configuration, which is characterized in that galactomannans is from golden cicada flower sporinite
It isolates and purifies to obtain, be made of mannose, glucose sugar, galactolipin and arabinose 1:5:2.5:1 polymerization in molar ratio, molecular weight
For 1.8kDa.
2. the galactomannans according to claim 1 rich in furans configuration, which is characterized in that the galactomannan is poly-
The strand connection type of sugar are as follows: → 2)-α-D-Manp (1 →, → 6)-α-D-Manp (1 →, → 4)-α-D-Glcp (1 →,
→ 2,4)-α-D-Glcp (1 →, → 5)-β-D-Galf (1 →, → 2)-β-D-Galf (1 →, molar ratio is followed successively by 1:1.3:
8:2:3:2。
3. a kind of preparation method of the galactomannans rich in furans configuration as claimed in claim 1 or 2, feature exist
In, comprising the following steps:
(1) by golden cicada flower sporinite grinding and sieving, acetone degreasing；
(2) the golden cicada flower sporinite after extracting degreasing extracts after adding water, is then centrifuged for obtaining supernatant；
(3) sporinite is spent as step (2) extract golden cicada repeatedly, and the supernatant merged after centrifugation obtains extracting solution；
(4) extracting solution obtained by step (3) is concentrated under reduced pressure, low-concentration ethanol solution precipitating is first added, after being centrifuged removal precipitating
High concentration ethanol solution precipitating is added, is centrifugally separating to obtain head product；
(5) head product after separation is dissolved with water, successively deproteination matter, dialysis desalination, are then freeze-dried to obtain raw sugar；
(6) raw sugar dissolution is placed on strong anion exchange column Q Sepharose TMFast Flow and is separated；
(7) using sodium chloride solution, successively it is maximum to choose absorbance for gradient elution, the simultaneously sulfuric acid-phynol method detection of collection eluent
Eluting peak where eluent be concentrated by evaporation, be dried to obtain final product.
4. the preparation method of the galactomannans according to claim 3 rich in furans configuration, which is characterized in that extraction
In the process, mass volume ratio 1g:20~30mL of golden cicada flower sporinite and water, 96~100 DEG C of extraction temperature, extraction time 4~
5h is extracted 2~5 times repeatedly.
5. the preparation method of the galactomannans according to claim 3 or 4 rich in furans configuration, which is characterized in that
The concentration of low-concentration ethanol solution is 40wt%~45wt% in step (4), and it is 4~6 times that leaching liquor is concentrated that volume, which is added, highly concentrated
Degree ethanol solution concentration is 77wt%~80wt%, and 4~6 times that volume is the solution after removal precipitating are added.
6. the preparation method of the galactomannans according to claim 3 or 4 rich in furans configuration, which is characterized in that
Sevag method deproteination matter is used in step (5), using the bag filter dialysis desalination of molecular cut off 3500Da.
7. the preparation method of the galactomannan sugar according to claim 3 or 4 rich in furans configuration, which is characterized in that step
Suddenly the separation process of (6) is as follows: the raw sugar product of 50mg being dissolved in the deionized water of 1~2mL, then 0.45 μm of microfiltration membranes
Filtering, then loading separation, loading speed are 1ml/min.
8. the preparation method of the galactomannans according to claim 3 rich in furans configuration, which is characterized in that step
(7) elution process is as follows: successively divide various concentration NaCl solution to gradient elution splitter to high concentration by from low concentration,
NaCl solution concentration gradient are as follows: 0,0.1 mol/L, 0.25 mol/L, 0.5 mol/L, elution flow rate 3mL/min, Ge Geti
Degree collects the eluent of 2~3 times of column volumes.
9. the preparation method of the galactomannans according to claim 3 or 8 rich in furans configuration, which is characterized in that
The wavelength of sulfuric acid-phynol method detection is 490nm.
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| CN104672339A (en) * | 2015-02-03 | 2015-06-03 | 浙江省林业科学研究院 | Cordyceps cicadae rhzomorph as well as preparation method and application thereof |
| CN107759707A (en) * | 2016-08-23 | 2018-03-06 | 浙江泛亚生物医药股份有限公司 | A kind of cicada fungus polysaccharide and its isolation and purification method |
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