CN109438531A - A kind of preparation method of high polymerization degree wood oligose monomer - Google Patents
A kind of preparation method of high polymerization degree wood oligose monomer Download PDFInfo
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- CN109438531A CN109438531A CN201811467966.0A CN201811467966A CN109438531A CN 109438531 A CN109438531 A CN 109438531A CN 201811467966 A CN201811467966 A CN 201811467966A CN 109438531 A CN109438531 A CN 109438531A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
- C07H3/06—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
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Abstract
The invention discloses a kind of preparation methods of high polymerization degree wood oligose monomer, belong to alga oligosaccharides extractive technique field.Wood oligose is prepared post separation high polymerization degree wood oligose monomer using Click Xlon using after the multiple precipitating of acetone by the present invention, realizes 4 sugar of wood for disposably preparing high-purity to wooden 8 sugar monomers, Xylotetrose is to the sugared purity of wood six all 95% or more.The present invention can disposably prepare the wood oligose monomer of high polymerization degree, especially realize 8 sugar monomer of wood for being difficult to separation and Extraction in the prior art；This method does not need secondarily purified, improves the yield of wood oligose monomer, reduces production cost；It is easy to operate, reproducible, application easy to spread.
Description
Technical field
The present invention relates to a kind of preparation methods of high polymerization degree wood oligose monomer, more particularly to one kind disposably to prepare height
The preparation method of the high polymerization degree wood oligose monomer of purity, belongs to alga oligosaccharides extractive technique field.
Background technique
Xylo-oligosaccharide is also known as wood oligose, is the functional polymerization being combined by 2-7 xylose molecule with β-Isosorbide-5-Nitrae glycosidic bond
Sugar.Compared with other oligosaccharide, the dosage for activating probiotics is small, and has heat-resisting, acidproof, reduction water activity and prevent from freezing
The various features such as knot, make it have wider application range.Wood oligose can be used as functional food additives, outstanding in food
It is widely applied in acid and fermented food；In medicine, wood oligose can be used for preventing and treating acute and chronic stomach
Inflammation, osteoporosis, cutaneous pruritus and tympanitis etc. maintain the retentiveness of excrement in addition, can increase intestines peristalsis,
Cardinal principle is the beneficial bacterium for activating enteron aisle；As feed addictive, using the forage feed livestock containing wood oligose and breed fish,
The immunity that livestock can be improved reduces various diseases, meanwhile, the growth cycle of livestock can be made to shorten；Agriculturally, wood oligose can
Growth stimulant and growth promoter as crops can be used as ripener to spray fruits and vegetables, and be remarkably improved
The yield of crops.
Wood oligose monomer can be used for wood oligose in medicine, food, aquaculture, modern agriculture, daily-use chemical industry and biochemical examination
The preparation research of reference substance, oligosaccharides chip that the exploitation of the fields such as agent new product, the check analysis of wood oligose and quality control；It is few
Carbohydrate chemistries and the glycobiology research such as sugar and the repercussion study of albumen, the structure-activity relationship of oligosaccharides drug and Study on mechanism
Field.The crucial technology of preparing of wood oligose monomer is the necessary condition of wood oligose wide popularization and application, therefore to wood oligose monomer
The research of technology of preparing has highly important scientific meaning.
In terms of wood oligose monomer preparation, the wood oligose of some low polymerization degrees is prepared using gel chromatographic columns or activated-charcoal column
There are reports for monomer, the components for xylose oligomer that a kind of degree of polymerization is 2~8 is disclosed such as patent 200510094118.6
Preparative separation method prepares low polymerization degree xylose monomers using ultrafiltration and multiple gel chromatography analysis；Patent
201410059775.6 disclose a kind of method for wood oligose being extracted from bamboo shoot shell and separation obtains wood oligose monomer, separation side
Method uses wood oligose monomer.Study on Preparation Technology at present about high polymerization degree wood oligose monomer is less, rarely seen high polymerization degree wood
Six sugar, wooden seven sugared, Xylooctaose the report, the technology for especially disposably preparing high polymerization degree wood oligose monomer, which has no, to be had been reported that.
Summary of the invention
In order to overcome the deficiencies in the prior art described above, the present invention provides a kind of preparation side of high polymerization degree wood oligose monomer
Method, realization disposably prepare high polymerization degree wood oligose monomer, especially prepare six sugar of wood, seven sugar of wood, Xylooctaose of high polymerization degree,
Fill up domestic technique blank.
The present invention is achieved through the following technical solutions above-mentioned technical effect:
A kind of preparation method of high polymerization degree wood oligose monomer, specifically includes the following steps:
1) prepared by high polymerization degree wood oligose: taking wood oligose 15-30g, the dissolution of 300ml distilled water is added, is made into 100-200mg/ml
Then solution adds 1000-1200ml acetone precipitating, high speed centrifugation 10-15min；The precipitating 150- that centrifugation is obtained
The dissolution of 200ml distilled water, adds 600-800ml acetone precipitating, high speed centrifugation 10-15min；Obtained precipitating will be centrifuged by, which continuing, uses
The dissolution of 80-100ml distilled water, adds 300-400ml acetone precipitating, high speed centrifugation 10-15min；The precipitating that finally centrifugation is obtained
It is dissolved out with 20-30ml distilled water, as high polymerization degree wood oligose；
2) Click Xlon prepares the filling of column: being seated in pillar after the methanol aqueous solution of Click Xlon filler is impregnated
In, column volume is 150-300ml after filling；
3) separation of high polymerization degree wood oligose monomer: taking and prepare column in the Click Xlon filled on the high poly- sample of appropriate wood,
With the same 5-8 column volume of flow velocity linear elution of methanol aqueous solution, elution fraction is collected, 4 sugar of wood, wood 5 can be obtained
Sugar, 6 sugar of wood, 7 sugar of wood and wooden 8 sugar monomers.
Above-mentioned high polymerization degree wood oligose is purchased from commercially available Shandong Long Li limited liability company, contains micro 8 sugar of wood.
Above-mentioned steps 1) in centrifugal rotational speed be 8000-12000r/min.
Above-mentioned steps 2) and 3) in methanol aqueous solution concentration be 50%-70%.
Above-mentioned steps 3) in methanol aqueous solution flow velocity be 1-2ml/min.
Preferably, a kind of preparation method of high polymerization degree wood oligose monomer, comprising the following steps:
1) prepared by high polymerization degree wood oligose: taking wood oligose 15-30g, the dissolution of 300ml distilled water is added, is made into 100-200mg/ml
Then solution adds 1000-1200ml acetone precipitating, high speed centrifugation 10-15min；The precipitating 150- that centrifugation is obtained
The dissolution of 200ml distilled water, adds 600-800ml acetone precipitating, high speed centrifugation 10-15min；Obtained precipitating will be centrifuged by, which continuing, uses
The dissolution of 80-100ml distilled water, adds 300-400ml acetone precipitating, high speed centrifugation 10-15min；The precipitating that finally centrifugation is obtained
It is dissolved out with 20-30ml distilled water, as high polymerization degree wood oligose；
2) Click Xlon prepares the filling of column: being seated in pillar after the methanol aqueous solution of Click Xlon filler is impregnated
In, column volume is 150-300ml after filling；
3) separation of high polymerization degree wood oligose monomer: taking and prepare column in the Click Xlon filled on the high poly- sample of appropriate wood,
With the same 5-8 column volume of flow velocity linear elution of methanol aqueous solution, elution fraction is collected, 4 sugar of wood, wood 5 can be obtained
Sugar, 6 sugar of wood, 7 sugar of wood and wooden 8 sugar monomers.
The present invention provides a kind of preparation method of high polymerization degree wood oligose monomer, compared with prior art, has following aobvious
Work advantage:
1) present invention passes through the ratio of rational proportion acetone and wood oligose, realizes the technology effect that can not be obtained in the prior art
Fruit realizes and disposably prepares the high polymerization degree wood oligose monomer of high-purity, Xylotetrose to six sugar purity of wood all 95% with
On, domestic blank has been filled up in the preparation of especially wooden 8 sugar monomers；
2) traditional wood oligose preparation method, needs multiple gel column to purify repeatedly, low efficiency；Test result shows general secondary
Purifying will lose 50% or more each time, and the present invention can disposably prepare the wood oligose monomer of high polymerization degree, not need two
Secondary purifying improves the yield of wood oligose monomer, reduces production cost；
3) operation of the present invention is simple, and disengaging time is short, reproducible, application easy to spread.
Detailed description of the invention
Fig. 1 is that the high polymerization degree wood oligose TLC of step 1) preparation detects figure；
Fig. 2 is that the TLC of step 3) eluent detects figure, and wherein 2-9 is 4 sugar of wood, and 28-40 is 5 sugar of wood, and 52-61 is 6 sugar of wood, 67-
73 be 7 sugar of wood, and 80-83 is 8 sugar of wood；
Fig. 3 is prepared by the present invention wooden 4 sugared HPLC chromatograms；
Fig. 4 is the wooden 4 sugar ESI-MS mass spectrogram prepared by the present invention；
Fig. 5 is prepared by the present invention wooden 5 sugared HPLC chromatograms；
Fig. 6 is the wooden 5 sugar ESI-MS mass spectrogram prepared by the present invention；
Fig. 7 is prepared by the present invention wooden 6 sugared HPLC chromatograms；
Fig. 8 is the wooden 6 sugar ESI-MS mass spectrogram prepared by the present invention；
Fig. 9 is prepared by the present invention wooden 7 sugared HPLC chromatograms；
Figure 10 is the wooden 7 sugar ESI-MS mass spectrogram prepared by the present invention；
Figure 11 is prepared by the present invention wooden 8 sugared HPLC chromatograms；
Figure 12 is the wooden 8 sugar ESI-MS mass spectrogram prepared by the present invention.
Specific embodiment
Embodiment 1
A kind of preparation method of high polymerization degree wood oligose monomer, comprising the following steps:
1) prepared by high polymerization degree wood oligose: taking wood oligose 20g, the dissolution of 300ml distilled water is added, is made into 150mg/ml solution, so
After add 1100ml acetone precipitating, 8000-12000r/min is centrifuged 10min；The precipitating 180ml distilled water that centrifugation is obtained
Dissolution, adds 700ml acetone precipitating, and 8000-12000r/min is centrifuged 10min；Continue to distill the precipitating that centrifugation obtains with 90ml
Water-soluble to add 350ml acetone precipitating out, 8000-12000r/min is centrifuged 10min；Finally the precipitating that centrifugation obtains is steamed with 25ml
Distilled water dissolution, as high polymerization degree wood oligose；
2) Click Xlon prepares the filling of column: after the methanol aqueous solution of Click Xlon filler 50%-70% concentration is impregnated
It is seated in pillar, column volume is 200ml after filling；
3) separation of high polymerization degree wood oligose monomer: taking and prepare column in the Click Xlon filled on the high poly- sample of appropriate wood,
With the same 5-8 column volume of flow velocity linear elution of the methanol aqueous solution of 50%-70% concentration, the flow velocity of methanol aqueous solution is 1-2ml/
Min is collected elution fraction, and 4 sugar of wood, 5 sugar of wood, 6 sugar of wood, 7 sugar of wood and wooden 8 sugar monomers can be obtained；
Above-mentioned high polymerization degree wood oligose is purchased from commercially available Shandong Long Li limited liability company.
Embodiment 2 detects high polymerization degree wood oligose using thin-layer chromatography (TLC)
Lamellae: German 60 F254 plate of Merck company's T LC Silica gel；
Solvent: n-butanol/formic acid/water；
Color developing agent: aniline/diphenylamines；
Test result is as shown in Figure 1；The high polymerization degree wood oligose obtained as seen from Figure 1 is mainly 4-8 sugar.
The eluent of 3 pairs of embodiment collections carries out thin-layer chromatography (TLC) analysis
Lamellae: German 60 F254 plate of Merck company's T LC Silica gel；
Solvent: n-butanol/formic acid/water；
Color developing agent: aniline/diphenylamines；
Test result is as shown in Figure 2；The high polymerization degree Xylotetrose that the present invention can disposably prepare high-purity as seen from Figure 2 arrives
Xylooctaose monomer.
The detection of 4 high polymerization degree wood oligose monomer of embodiment
(1) High Performance Gel Permeation chromatography (HPGPC) is analyzed
Detecting instrument: Japanese Shimadzu liquid chromatograph；Chromatographic column: Click Xion (250mm × 4.6 mm, 10 μm)；Flowing
Phase: A water, B methanol；Detector: evaporative light scattering detector；
(2) electrospray ionization mass spectrum (ESI-MS) is analyzed
Detecting instrument: U.S.'s Micromass Q-TOF Ultima Global Quadrupole-time of flight mass spectrometry instrument；Mass spectrum inspection
Survey condition: anionic textiles mode；
(3) for testing result as shown in Fig. 3-12, test result shows that the present invention may be implemented to prepare 4 sugar of wood, the wood 5 of high-purity
Sugar, 6 sugar of wood, 7 sugar of wood and wooden 8 sugar monomers.
The above examples are only used to illustrate the technical scheme of the present invention, rather than is limited；Although referring to aforementioned reality
It applies example to be described in detail to by invention, but for those of ordinary skill in the art, it still can be to aforementioned reality
Technical solution documented by example is applied to modify or equivalent replacement of some of the technical features；And to these modifications
Or replacement, the spirit and scope for claimed technical solution of the invention that it does not separate the essence of the corresponding technical solution.
Claims (5)
1. a kind of preparation method of high polymerization degree wood oligose monomer, it is characterised in that the preparation method specifically includes following step
It is rapid:
1) prepared by high polymerization degree wood oligose: taking wood oligose 15-30g, the dissolution of 300ml distilled water is added, is made into 100-200mg/ml
Then solution adds 1000-1200ml acetone precipitating, high speed centrifugation 10-15min；The precipitating 150- that centrifugation is obtained
The dissolution of 200ml distilled water, adds 600-800ml acetone precipitating, high speed centrifugation 10-15min；Obtained precipitating will be centrifuged by, which continuing, uses
The dissolution of 80-100ml distilled water, adds 300-400ml acetone precipitating, high speed centrifugation 10-15min；The precipitating that finally centrifugation is obtained
It is dissolved out with 20-30ml distilled water, as high polymerization degree wood oligose；
2) Click Xlon prepares the filling of column: being seated in pillar after the methanol aqueous solution of Click Xlon filler is impregnated
In, column volume is 150-300ml after filling；
3) separation of high polymerization degree wood oligose monomer: taking and prepare column in the Click Xlon filled on the high poly- sample of appropriate wood,
With the same 5-8 column volume of flow velocity linear elution of methanol aqueous solution, elution fraction is collected, 4 sugar of wood, wood 5 can be obtained
Sugar, 6 sugar of wood, 7 sugar of wood and wooden 8 sugar monomers.
2. a kind of preparation method of high polymerization degree wood oligose monomer according to claim 1, it is characterised in that the step
1) centrifugal rotational speed is 8000-12000r/min in.
3. a kind of preparation method of high polymerization degree wood oligose monomer according to claim 1, it is characterised in that the step
And 3) 2) concentration of methanol aqueous solution is 50%-70% in.
4. a kind of preparation method of high polymerization degree wood oligose monomer according to claim 1, it is characterised in that the step
3) flow velocity of methanol aqueous solution is 1-2ml/min in.
5. a kind of preparation method of high polymerization degree wood oligose monomer according to claim 1, it is characterised in that the preparation
Method preferably includes following steps:
1) prepared by high polymerization degree wood oligose: taking wood oligose 15-30g, the dissolution of 300ml distilled water is added, is made into 100-200mg/ml
Then solution adds 1000-1200ml acetone precipitating, high speed centrifugation 10-15min；The precipitating 150- that centrifugation is obtained
The dissolution of 200ml distilled water, adds 600-800ml acetone precipitating, high speed centrifugation 10-15min；Obtained precipitating will be centrifuged by, which continuing, uses
The dissolution of 80-100ml distilled water, adds 300-400ml acetone precipitating, high speed centrifugation 10-15min；The precipitating that finally centrifugation is obtained
It is dissolved out with 20-30ml distilled water, as high polymerization degree wood oligose；
2) Click Xlon prepares the filling of column: being seated in pillar after the methanol aqueous solution of Click Xlon filler is impregnated
In, column volume is 150-300ml after filling；
3) separation of high polymerization degree wood oligose monomer: taking and prepare column in the Click Xlon filled on the high poly- sample of appropriate wood,
With the same 5-8 column volume of flow velocity linear elution of methanol aqueous solution, elution fraction is collected, 4 sugar of wood, wood 5 can be obtained
Sugar, 6 sugar of wood, 7 sugar of wood and wooden 8 sugar monomers.
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| CN201811467966.0A CN109438531A (en) | 2018-12-03 | 2018-12-03 | A kind of preparation method of high polymerization degree wood oligose monomer |
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| CN201811467966.0A CN109438531A (en) | 2018-12-03 | 2018-12-03 | A kind of preparation method of high polymerization degree wood oligose monomer |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1847254A (en) * | 2005-08-30 | 2006-10-18 | 南京林业大学 | Prepn and separation process of components for xylose oligomer of polymerization degree 2-8 |
| CN103789376A (en) * | 2014-02-21 | 2014-05-14 | 青岛博智汇力生物科技有限公司 | Method of extracting xylo-oligosaccharide from bamboo shoot shells and separating to obtain xylo-oligosaccharide monomer |
| CN104164520A (en) * | 2014-07-15 | 2014-11-26 | 南京林业大学 | Method using acetic acid for controlled catalysis of orientational hydrolysis of xylan to prepare multicomponent xylooligosaccharide |
-
2018
- 2018-12-03 CN CN201811467966.0A patent/CN109438531A/en not_active Withdrawn
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1847254A (en) * | 2005-08-30 | 2006-10-18 | 南京林业大学 | Prepn and separation process of components for xylose oligomer of polymerization degree 2-8 |
| CN103789376A (en) * | 2014-02-21 | 2014-05-14 | 青岛博智汇力生物科技有限公司 | Method of extracting xylo-oligosaccharide from bamboo shoot shells and separating to obtain xylo-oligosaccharide monomer |
| CN104164520A (en) * | 2014-07-15 | 2014-11-26 | 南京林业大学 | Method using acetic acid for controlled catalysis of orientational hydrolysis of xylan to prepare multicomponent xylooligosaccharide |
Non-Patent Citations (1)
| Title |
|---|
| MING-HSU CHEN，等: "Autohydrolysis of Miscanthus x giganteus for the production of xylooligosaccharides (XOS): Kinetics, characterization and recovery", 《BIORESOURCE TECHNOLOGY》 * |
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