CN109942723A - A kind of separation method of polysaccharides - Google Patents
A kind of separation method of polysaccharides Download PDFInfo
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Abstract
The invention discloses a kind of separation method of polysaccharides, include the following steps: that (1) chooses Lycium fruit, through water heating extracting 2~3 times, extracting solution merges, and concentration adds ethyl alcohol to make to precipitate, and collection precipitating is simultaneously dry, obtains fructus lycii total starches；(2) standby liquid phase systems are suppressed in the use of the fructus lycii total starches obtained by step (1) to be purified, first gradient elution is carried out as eluant, eluent through weak anion exchange column chromatography, buffer, polysaccharides component efflux needed for being collected by chromatographic peak, it is concentrated again through ultrafiltration membrane, molecular exclusion column chromatography desalination, it is dry, obtain the polysaccharides that purity is greater than 80%.The present invention can greatly improve purification process development efficiency, effectively promotion separative efficiency, optimize separating effect, remove a large amount of impurity such as pigment, the metal ion in total starches, achieve the purpose that improve column effect, favorable reproducibility, be easy amplification preparative-scale.
Description
Technical field
The present invention relates to a kind of separation methods of polysaccharides, belong to technical field of biochemical industry.
Background technique
About 80 kinds of the Solanaceae Lycium whole world is distributed mainly on South America zone of constant temperature area.There are lycium barbarum, north in China
Square fructus lycii (Hebei), red branch fructus lycii (Xinjiang), cuts calyx fructus lycii (Inner Mongol), column casing fructus lycii (Xinjiang), Yunnan fructus lycii at Lycium barbarum
The kinds such as (Yunnan), black fruit fructus lycii (Qinghai), yellow fruit fructus lycii, wherein lycium barbarum is to obtain scaled artificial planting only at present
One kind is the mainstream product in the market.Fructus lycii is the dual-purpose of drug and food functional food of Chinese tradition, and it is Ningxia that Chinese Pharmacopoeia, which records fructus lycii,
The dry mature fruit of fructus lycii (Lycium barbarum), function tonification liver kidney, benefiting shrewd head are used for consumption consumptive loss, waist and knee acid
Bitterly, dizziness and tinnitus, impotence and seminal emission, Heat Diabetes, blood deficiency chlorosis, blurred vision.In fructus lycii containing polysaccharides, luteole,
Glycine betaine, flavones and other functional ingredient, these functional components, which have, adjusts immune, anti-inflammatory, protection cell, blood vessel, nerve, Yi Jikang
Aging and the multiple pharmacological effects such as antitumor.Modern pharmacological research proves that main ingredient polysaccharides contained by fructus lycii is that it is most heavy
One of the effect of wanting ingredient, it is a kind of complicated water-soluble sugar albumen, immunological regulation, it is hypoglycemic the effects of it is more bright
Really.Fructus lycii started to export to Europe, North America etc. all over the world at the beginning of 21 century, was known as super fruit, and health-care efficacy obtains
To international endorsement.In recent years, the development and application of Lycium barbarum polysaccharide extract are favored by market.
The traditional handicraft relevant report for extracting polysaccharides is more.It is conventional to use simple water extract-alcohol precipitation method, it can also tie
The methods of ultrafiltration, ultrasonic extraction is closed to carry out.But since polysaccharides relative amount is lower, complicated composition, very using traditional handicraft
Difficulty isolates and purifies it, and general polysaccharides content is below 60%." fermentation method mentions patent 2005100467857
Take polysaccharides ", a kind of method that applied bioengineering technology extracts polysaccharides is disclosed, it was grown using Angel Yeast
Monosaccharide and disaccharide is consumed in journey, the means of nitrogen substance improve the purity of polysaccharides, entire technical process include crush, leaching
Easily there is fermentation not exclusively in this method production process and impurity are more in bubble, aerobic fermentation, filtering, decoloration, removal of impurities, vacuum distillation
The problems such as；Patent 021153825 " a kind of polysaccharides and its preparation method and application " discloses a kind of using membrane separation technique
The method for extracting polysaccharides, entire technical process are dense including shearing, squeezing, centrifugation, hollow-fibre membrane filtering, hollow-fibre membrane
Contracting, freeze-drying and etc.；Patent 201610762613.8 " a kind of preparation method of polysaccharides ", also discloses polysaccharides
Technique, including screening and removing impurities, drying, crushing, separation, supercritical CO2Extraction, immersion, enzymatic hydrolysis, centrifugation, desalination, level Four are super
Then filter, reverse osmosis concentration, drying bu sublimation, low-temperature grinding are packaged as finished product；Above method complex process, production
Yield is low.The laboratory purifying process of polysaccharides generally exchanges pillar layer separation, each flow point warp of collection using atmospheric pressure ion
Offline ultraviolet detection analyzes its purity, merges the flow point of identical chromatographic peak, then continue to purify through subsequent technique, low separation efficiency
Under, it is difficult to realize the prepare with scale of high-purity polysaccharides.
Summary of the invention
Goal of the invention of the invention is to provide a kind of separation method of polysaccharides, and separation process can be monitored online, and reach
To separating degree height, favorable reproducibility, the purpose for being easy amplification preparative-scale, so as to prepare the fructus lycii that purity is greater than 80%
Polysaccharide, and greatly improve separative efficiency.
To achieve the above object of the invention, the technical solution adopted by the present invention is that: a kind of separation method of polysaccharides, including
Following steps: (1) choosing Lycium fruit, and through water heating extracting 2~3 times, extracting solution merges, and concentration adds ethyl alcohol to make to sink
It forms sediment, collects precipitating and dry, acquisition fructus lycii total starches；
(2) standby liquid phase systems are suppressed in the use of the fructus lycii total starches obtained by step (1) to be purified, first hand over through weak anionic
Column chromatography, buffer are changed as eluant, eluent and carries out gradient elution, polysaccharides component efflux needed for being collected by chromatographic peak, then pass through
Ultrafiltration membrane concentration, molecular exclusion column chromatography desalination is dry, obtains the polysaccharides that purity is greater than 80%.
In above-mentioned technical proposal, in the step (1), Lycium can be selected from lycium barbarum, northern Lycium chinense (Hebei),
Lycium barbarum, cuts calyx fructus lycii (Inner Mongol), column casing fructus lycii (Xinjiang), Yunnan fructus lycii (Yunnan), black fruit fructus lycii at red branch fructus lycii (Xinjiang)
The kinds such as (Qinghai), yellow fruit fructus lycii, it is conventional to select lycium barbarum.The fruit is dry fruit or fresh fruit.Polysaccharide generally uses heat
Water refluxing extraction, the dregs of a decoction are used further to extract after wherein medlar fresh fruit directly can be extracted or be squeezed the juice；Dried fruit of lycium barbarum can be after water infiltration
It extracts again.Result of study of the present invention show polysaccharides molecular weight generally 60,000 or more, thus extracting solution condensing mode
It is preferred that ultrafiltration membrane is concentrated, the hollow-fibre membrane concentration of more preferable 3~10kD of molecular cut off.Concentrate can add 2 times of amount ethyl alcohol to make
Precipitating collects precipitating, vacuum dried, spray drying or freeze-drying, obtains fructus lycii total starches.
Standby liquid phase systems are suppressed in using in above-mentioned technical proposal, in the step (2) to purify fructus lycii total starches.
Medium pressure prepare liquid phase systems be equipped with conductance/pH/DAD Series detectors, so as to realize to salinity (Conductivity detection),
The synchronization on-line checking of pH value, desired polysaccharide sample chromatogram peak (DAD, feature multi-wavelength detection) realizes process parameter optimizing, color
Compose the Intelligent information acquisition of separating effect analysis and target component collection.For polysaccharides, can select ultraviolet 210nm and
280nm monitors separating effect.
In above-mentioned technical proposal, separated in the step (2) using weak anion exchange column, wherein it is preferred that DEAE-650M
Filler can effectively remove the impurity components such as the pigment in fructus lycii total starches.The equilibrium liquid type of the buffer can be selected more
Kind inorganic salt solution, wherein it is preferred that Tris (trishydroxymethylaminomethane)-HCl, more preferable 30-50mM Tris-HCl；Most preferably
40mM Tris-HCl(pH7.0-9.5).The preferred 0.3-0.5M NaCl of high salt eluent type of the buffer (contains 40mM
Tris-HCl, pH7.0-9.5), the preferred linear gradient elution of gradient elution mode, elution volume can according to need choosing
It selects, wherein it is preferred that 10 times of column volumes are eluted.The eluent collection mode of polysaccharides target components, is received online using by peak
Integrated mode.
In above-mentioned technical proposal, polysaccharides component efflux is concentrated through ultrafiltration membrane again in the step (2), wherein it is preferred that
The hollow-fibre membrane of 3~10kD of molecular cut off.Concentrate again through molecular exclusion column chromatography desalination, preferably Sephadex filler and
It is middle to suppress standby liquid phase systems, on-line monitoring desalination is carried out by mobile phase of pure water.The polysaccharides flow point collected after desalination, warp
Freeze-drying can get the polysaccharides product that purity is greater than 80%.
Due to the above technical solutions, the present invention has the following advantages over the prior art:
The present invention can greatly improve purification process development efficiency, effectively promotion separative efficiency, optimize separating effect, improve
Polysaccharides extraction process stability removes a large amount of impurity such as pigment, the metal ion in total starches, reaches raising column and imitates, again
Existing property is good, is easy the purpose of amplification preparative-scale.
Detailed description of the invention
Fig. 1 is the middle compacting of fructus lycii total starches described in the embodiment of the present invention 1 for liquid phase systems DEAE-650M chromatography post separation
Chromatogram, ultraviolet detection wavelength are 210nm and 280nm.In figure, P1-P4 is the polysaccharides sample chromatogram peak collected by peak.
Fig. 2 is the efficient molecular-exclusion chromatography figure of polysaccharides sample P 2 described in the embodiment of the present invention 1.
Fig. 3 is the efficient molecular-exclusion chromatography figure of polysaccharides sample P 3 described in the embodiment of the present invention 1.
Specific embodiment
The invention will be further described with reference to the accompanying drawings and embodiments:
Embodiment one:
Referring to shown in Fig. 1 to 3, a kind of separation method of polysaccharides includes the following steps:
(1) Lycium fruit is chosen, through water heating extracting 2~3 times, extracting solution merges, and concentration adds ethyl alcohol to make to sink
It forms sediment, collects precipitating and dry, acquisition fructus lycii total starches；
(2) standby liquid phase systems are suppressed in the use of the fructus lycii total starches obtained by step (1) to be purified, first hand over through weak anionic
Column chromatography, buffer are changed as eluant, eluent and carries out gradient elution, polysaccharides component efflux needed for being collected by chromatographic peak, then pass through
Ultrafiltration membrane concentration, molecular exclusion column chromatography desalination is dry, obtains the polysaccharides that purity is greater than 80%.
In the present embodiment, in the step (1), Lycium can be selected from lycium barbarum, northern Lycium chinense (Hebei), Xinjiang
Fructus lycii, red branch fructus lycii (Xinjiang) cut calyx fructus lycii (Inner Mongol), column casing fructus lycii (Xinjiang), Yunnan fructus lycii (Yunnan), black fruit fructus lycii (blueness
Sea), the kinds such as yellow fruit fructus lycii, it is conventional to select lycium barbarum.The fruit is dry fruit.Polysaccharide generally uses hot water return to extract,
The dregs of a decoction are used further to extract after wherein medlar fresh fruit directly can be extracted or be squeezed the juice；Dried fruit of lycium barbarum can extract again after water infiltration.
According to polysaccharides molecular weight generally 60,000 or more, extracting solution condensing mode selects ultrafiltration membrane concentration.Concentrate can add 2 times of amounts
Ethyl alcohol makes to precipitate, and collects precipitating, vacuum dried, spray drying or freeze-drying, obtains fructus lycii total starches.
Standby liquid phase systems are suppressed in using in the present embodiment, in the step (2) to purify fructus lycii total starches.It is described
It is middle to suppress standby liquid phase systems outfit conductance/pH/DAD Series detectors, so as to realize to salinity (Conductivity detection), pH
The synchronization on-line checking of value, desired polysaccharide sample chromatogram peak (DAD, feature multi-wavelength detection) realizes process parameter optimizing, chromatography
The Intelligent information acquisition that separating effect analysis and target component are collected.For polysaccharides, can select ultraviolet 210nm and
280nm monitors separating effect.
It in the present embodiment, is separated in the step (2) using weak anion exchange column, selects DEAE-650M filler, it can
Effectively to remove the impurity components such as the pigment in fructus lycii total starches.Tris (three hydroxyls can be selected in the equilibrium liquid type of the buffer
Aminomethane)-HCl.The high salt eluent type of the buffer select 0.3M NaCl (Tris-HCl containing 40mM,
PH7.0-9.5), the gradient elution mode selects linear gradient elution, and elution volume can according to need selection, select here
10 times of column volumes are eluted.The eluent collection mode of polysaccharides target components, using by the online collection mode in peak.
In the present embodiment, polysaccharides component efflux is concentrated through ultrafiltration membrane again in the step (2), selects retention here
The hollow-fibre membrane of 3~10kD of molecular weight.Concentrate through molecular exclusion column chromatography desalination, selects Sephadex filler and middle pressure again
Liquid phase systems are prepared, carry out on-line monitoring desalination by mobile phase of pure water.The polysaccharides flow point collected after desalination, it is chilled
It is dry, it can get the polysaccharides product that purity is greater than 80%.
Embodiment two:
A kind of separation method of polysaccharides, includes the following steps:
(1) Lycium fruit is chosen, through water heating extracting 2~3 times, extracting solution merges, and concentration adds ethyl alcohol to make to sink
It forms sediment, collects precipitating and dry, acquisition fructus lycii total starches；
(2) standby liquid phase systems are suppressed in the use of the fructus lycii total starches obtained by step (1) to be purified, first hand over through weak anionic
Column chromatography, buffer are changed as eluant, eluent and carries out gradient elution, polysaccharides component efflux needed for being collected by chromatographic peak, then pass through
Ultrafiltration membrane concentration, molecular exclusion column chromatography desalination is dry, obtains the polysaccharides that purity is greater than 80%.
In the present embodiment, in the step (1), Lycium can be selected from lycium barbarum, northern Lycium chinense (Hebei), Xinjiang
Fructus lycii, red branch fructus lycii (Xinjiang) cut calyx fructus lycii (Inner Mongol), column casing fructus lycii (Xinjiang), Yunnan fructus lycii (Yunnan), black fruit fructus lycii (blueness
Sea), the kinds such as yellow fruit fructus lycii, it is conventional to select lycium barbarum.The fruit is fresh fruit.Polysaccharide generally uses hot water return to extract,
The dregs of a decoction are used further to extract after wherein medlar fresh fruit directly can be extracted or be squeezed the juice；Dried fruit of lycium barbarum can extract again after water infiltration.
According to polysaccharides molecular weight generally 60,000 or more, extracting solution condensing mode selects the doughnut of 3~10kD of molecular cut off
Film concentration.Concentrate can add 2 times of amount ethyl alcohol to make to precipitate, and collect precipitating, vacuum dried, spray drying or freeze-drying, obtain
Obtain fructus lycii total starches.
It in the present embodiment, is separated in the step (2) using weak anion exchange column, selects DEAE-650M filler, it can
Effectively to remove the impurity components such as the pigment in fructus lycii total starches.40mM Tris- can be selected in the equilibrium liquid type of the buffer
HCl.The high salt eluent type of the buffer selects 0.3M NaCl (Tris-HCl containing 40mM, pH7.0-9.5), the ladder
It spends type of elution and selects linear gradient elution, elution volume can according to need selection, select 10 times of column volumes to be washed here
It is de-.The eluent collection mode of polysaccharides target components, using by the online collection mode in peak.
Embodiment three:
A kind of separation method of polysaccharides, includes the following steps:
(1) Lycium fruit is chosen, through water heating extracting 2~3 times, extracting solution merges, and concentration adds ethyl alcohol to make to sink
It forms sediment, collects precipitating and dry, acquisition fructus lycii total starches；
(2) standby liquid phase systems are suppressed in the use of the fructus lycii total starches obtained by step (1) to be purified, first hand over through weak anionic
Column chromatography, buffer are changed as eluant, eluent and carries out gradient elution, polysaccharides component efflux needed for being collected by chromatographic peak, then pass through
Ultrafiltration membrane concentration, molecular exclusion column chromatography desalination is dry, obtains the polysaccharides that purity is greater than 80%.
It in the present embodiment, is separated in the step (2) using weak anion exchange column, selects DEAE-650M filler, it can
Effectively to remove the impurity components such as the pigment in fructus lycii total starches.40mM Tris- can be selected in the equilibrium liquid type of the buffer
HCl(pH7.0-9.5).The high salt eluent type of the buffer selects 0.3M NaCl (Tris-HCl containing 40mM, pH7.0-
9.5), the gradient elution mode selects linear gradient elution, and elution volume can according to need selection, selects 10 times of columns here
Volume is eluted.The eluent collection mode of polysaccharides target components, using by the online collection mode in peak.
Binding ability, resolution ratio and the selective property for depending not only on filler and ligand and aglucon of weak anionic
Density, additionally depends on specific experiment condition such as pH and elution requirement, thus need to pH to the mobile phase employed in separation process, from
Sub- intensity and elution requirement are investigated.Preparative liquid chromatograph (Li Sui Science and Technology Ltd.) is pressed using in APPS MV 10D,
Chromatographic column 2.5 × 20cm of specification (column volume 100mL) carries out following single factor exploration by index of the separating effect of polysaccharides
Experiment.
(1) investigation of the different fillers to polysaccharides separating effect
The present invention has primarily looked at following influence of several fillers to the separating effect of polysaccharides: strong anion exchange is filled out
Expect (GP-Q30), hydrophobic chromatography filler (SP), weak anionic displacement chromatography filler (DEAE80, DEAE-650M, DEAE-FF).It surveys
Test result shows: strong anion exchange filler, hydrophobic chromatography filler cannot separate polysaccharides well；Weak anionic exchange
Chromatographic stuffing is substantially better than other type fillers to the separating property of polysaccharides, but different manufacturers and the weak yin of different model from
Sub- displacement chromatography filler separating capacity also has larger gap, preferable through investigating DEAE-650M, DEAE-FF effect.
(2) investigation of buffer type and pH
Respectively with deionized water (pH8.0), 40mM sodium chloride (pH8.0), 40mM ammonium sulfate (pH8.0), 40mM Tris-
HCl (pH8.0) is mobile phase A (balance phase).Through investigating, in addition to water, other buffers have certain separating effect；Wherein with
Tris-HCl (pH8.0) separating degree is more excellent.
The pH working range of DEAE filler is 3-10, but available data has no the correlative study of polysaccharides separating effect.
The present invention selects 40mM Tris-HCl, investigates different pH (4.0,6.0,7.0,7.5,8.0,8.5,9.0,9.5,10.0) respectively
For the separating effect of mobile phase A.As a result it has been surprisingly found that, different pH have a significant impact separating degree, and in pH7.0 or less, fructus lycii is more
Sugar is difficult to separate, only best to the separating effect of polysaccharides in pH7.5-9.5 range.
Investigate influence of the various concentration buffer to the separating effect of polysaccharides.Respectively with 10,20,30,40,50mM
Tris-HCl (pH9.0) is that initial liquid phase is investigated.The result shows that sample starts to elute in conductance 0.64ms/cm, it should
It is preferred that the correspondence buffer concentration 40mM under conductance.
(3) investigation of elution requirement
Through high salt concentration eluent (Mobile phase B) investigate, determine with 0.3-0.5M NaCl (Tris-HCl containing 40mM,
PH9.0) more excellent, wherein 0.3M NaCl separating degree is best.It is investigated by elution volume (5,10,15,20BV), elution volume is
Separating degree can be met the requirements when 10BV.
In the selection of separation method, there are stepwise elution and two kinds of linear gradient elution, stepwise elution operationally compares
Simply, but in ionic strength acute variation the substance summit eluted be easy to cause stepwise elution very close to being even overlapped
Starting includes multiple components in the peak to get off, and linear elution is easy preparation, and easily repeats, and therefore, this experiment uses line
The clastotype of property ionic strength gradient elution.
(4) investigation of applied sample amount and flow velocity
When investigation applied sample amount is respectively 0.3,0.5,0.7g, the combination and separating capacity of filler.As a result, it has been found that DEAE-
Every milliliter of filler combination 3-5mg sample of the best volume containing the sample average out to of 650M, therefore applied sample amount of the present invention is set to 0.5g (column volume
100mL)。
Polysaccharides separating effect when flow velocity is 3,5,7ml/min is investigated respectively, in conjunction with applied sample amount and elution volume, separation
When working flow rate be set to 5mL/mL, balance is set to flow velocity when cleaning splitter: 8ml/min (98cm/h).
(5) preferable separate condition
By the studies above, Optimizing Technical of the present invention is selected are as follows:
(1) lycium barbarum fruit is chosen, through water heating extracting 2~3 times, extracting solution merges, through molecular cut off 10kD's
Hollow-fibre membrane concentration, adds 2 times of ethyl alcohol to make to precipitate, and collects and precipitates and be freeze-dried, and obtains fructus lycii total starches；
(2) it suppresses standby liquid phase systems in the use of the fructus lycii total starches obtained by (1) to be purified, first through the weak yin of DEAE-650M
Ion exchange column chromatography, 40mM Tris-HCl (pH9.0) and 0.3M NaCl (Tris-HCl containing 40mM, pH9.0) are as elution
Agent carries out linear gradient elution, sample detection wavelength 210 and 280nm, the stream of polysaccharides component P1-P4 needed for collecting by chromatographic peak
Liquid out, then the concentration of the hollow-fibre membrane ultrafiltration membrane through molecular cut off 10kD, Sephadex G25 molecular exclusion column chromatography desalination,
Freeze-drying obtains the polysaccharides P1-P4 sample that purity is greater than 80%.The middle compacting of fructus lycii total starches is for liquid phase systems
DEAE-650M post separation chromatogram is shown in Fig. 1.
(6) polysaccharides product testing
Using efficient molecular-exclusion chromatography chromatography, testing product peak purity and molecular weight.Chromatographic condition: TSK
G4000PWXLChromatographic column (7.8 × 30cm)；Mobile phase 0.2M NaCl；Flow velocity 0.6mLmin-1；30 DEG C of column temperature；Ultraviolet detection
Wavelength 210nm；20 μ L of sample volume.Use the glucan (5.0mg/mL) of T10, T40, T70, T110, T200 Series Molecules amount for
Standard items measure the relative molecular weight of polysaccharides.The compound concentration 1.0mg/mL of polysaccharides test sample.
Through testing and analyzing, the results showed that, one group polysaccharide of the polysaccharides mainly by molecular weight greater than 60,000 forms, these are more
Glycan molecule amount is close, but the different present invention of institute's band protein content select DEAE-650M weak anion exchange column chromatography, can get
Preferably separation.Wherein, polysaccharides P1-P4 is all larger than 80% through detecting peak purity；P2, P3 are that purity is uniform greater than 90%
Polysaccharide, testing result are shown in Fig. 2 and Fig. 3 respectively.
The foregoing description of the disclosed embodiments enables those skilled in the art to implement or use the present invention.
Various modifications to these embodiments will be readily apparent to those skilled in the art, as defined herein
General Principle can be realized in other embodiments without departing from the spirit or scope of the present invention.Therefore, of the invention
It is not intended to be limited to the embodiments shown herein, and is to fit to and the principles and novel features disclosed herein phase one
The widest scope of cause.
Claims (9)
1. a kind of separation method of polysaccharides, which comprises the steps of:
(1) Lycium fruit is chosen, through water heating extracting 2~3 times, extracting solution merges, and concentration adds ethyl alcohol to make to precipitate, and receives
Collection precipitates and dry, acquisition fructus lycii total starches；
(2) it suppresses standby liquid phase systems in the use of the fructus lycii total starches obtained by step (1) to be purified, first through weak anion exchange column
Chromatography, buffer carry out gradient elution as eluant, eluent, polysaccharides component efflux needed for collecting by chromatographic peak, then through ultrafiltration
Film concentration, molecular exclusion column chromatography desalination is dry, obtains the polysaccharides that purity is greater than 80%.
2. the separation method of polysaccharides according to claim 1, it is characterised in that: Lycium is planted in the step (1)
Object fruit selects dry fruit or fresh fruit.
3. the separation method of polysaccharides according to claim 1, it is characterised in that: Lycium is planted in the step (1)
Object fruit selects lycium barbarum.
4. the separation method of polysaccharides according to claim 1, it is characterised in that: extracting solution is dense in the step (1)
Contracting mode selects ultrafiltration membrane concentration or the hollow-fibre membrane concentration of 3~10kD of molecular cut off.
5. the separation method of polysaccharides according to claim 1, it is characterised in that: the middle compacting in the step (2)
Standby liquid phase systems are equipped with conductance/pH/DAD Series detectors.
6. the separation method of polysaccharides according to claim 1, it is characterised in that: in the step (2) it is weak yin from
Sub- exchange column selects DEAE-650M filler.
7. the separation method of polysaccharides according to claim 1, it is characterised in that: the equilibrium liquid type of the buffer
Selecting Tris-HCl 30-50mM Tris-HCl or 40mM Tris-HCl, the pH of the 40mM Tris-HCl is 7.0-
9.5。
8. the separation method of polysaccharides according to claim 1, it is characterised in that: the high salt eluent of the buffer
Type selects 0.3-0.5M NaCl, contains 40mM Tris-HCl, the pH7.0-9.5 of the 40mM Tris-HCl, the gradient
Type of elution selects linear gradient elution.
9. the separation method of polysaccharides according to claim 1, it is characterised in that: the ultrafiltration membrane in the step (2)
The hollow-fibre membrane of 3~10kD of molecular cut off is selected, the molecular exclusion column chromatography selects Sephadex filler.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110746515A (en) * | 2019-10-28 | 2020-02-04 | 百瑞源枸杞股份有限公司 | Lycium barbarum polysaccharide, lycium barbarum red element and lycium barbarum polypeptide prepared by synchronous separation and preparation method thereof |
| CN112521524A (en) * | 2020-12-24 | 2021-03-19 | 中国科学院兰州化学物理研究所 | Composition for synergistically reducing blood sugar by using lycium barbarum polysaccharide and acaudina molpadioides functional peptide |
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| CN112521524A (en) * | 2020-12-24 | 2021-03-19 | 中国科学院兰州化学物理研究所 | Composition for synergistically reducing blood sugar by using lycium barbarum polysaccharide and acaudina molpadioides functional peptide |
| CN112521524B (en) * | 2020-12-24 | 2022-05-24 | 中国科学院兰州化学物理研究所 | Composition for synergistically reducing blood sugar by using lycium barbarum polysaccharide and acaudina molpadioides functional peptide |
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