CN108912412A - A kind of preparation method of agar polysaccharide magnetic microsphere - Google Patents
A kind of preparation method of agar polysaccharide magnetic microsphere Download PDFInfo
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- CN108912412A CN108912412A CN201810872992.5A CN201810872992A CN108912412A CN 108912412 A CN108912412 A CN 108912412A CN 201810872992 A CN201810872992 A CN 201810872992A CN 108912412 A CN108912412 A CN 108912412A
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/12—Powdering or granulating
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
- C08J2305/12—Agar-agar; Derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K3/00—Use of inorganic substances as compounding ingredients
- C08K3/18—Oxygen-containing compounds, e.g. metal carbonyls
- C08K3/20—Oxides; Hydroxides
- C08K3/22—Oxides; Hydroxides of metals
- C08K2003/2265—Oxides; Hydroxides of metals of iron
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K2201/00—Specific properties of additives
- C08K2201/01—Magnetic additives
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
- C08K2201/00—Specific properties of additives
- C08K2201/011—Nanostructured additives
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Abstract
The present invention relates to a kind of preparation methods of agar polysaccharide magnetic microsphere, belong to separation and purification of biological macromolecule technical field.The present invention prepares magnetic iron oxide powder by raw material of frerrous chloride,During preparation,It joined methanol into pre-reaction liquid,Using methanol as raw material,So that methanol and ammonia reaction generate organic amine,And the polar group being rich in organic amine has certain electron supplying capacity,Iron ion and ferrous ion surface form one layer of monomolecular film in pre-reaction liquid,Prevent magnetic iron oxide particles partial size excessive,The ammonia that the present invention is passed through, which is dissolved in after water, to be obtained ammonium hydroxide and can be used as coprecipitator,It is small that partial size is made,The nano-scale magnetic microballoon of size uniformity,Finally by inverse phase suspension,Pass through hydrogen bond using agarose,Coordinate bond absorption is wrapped on nano-scale magnetic microballoon crystal grain,Form close chondritic,It is finally made agar polysaccharide magnetic microsphere,Agar polysaccharide magnetic microsphere partial size produced by the present invention is small,It is evenly distributed,It has broad application prospects.
Description
Technical field
The present invention relates to a kind of preparation methods of agar polysaccharide magnetic microsphere, belong to separation and purification of biological macromolecule technology neck
Domain.
Background technique
Agarose is linear polymer, and basic structure is in the β-D- galactolipin of 1,3 connections and 3,6- of Isosorbide-5-Nitrae connection
The long-chain that ether-L- galactolipin alternately connects.Agar pectin is by the heterogeneous mixture of many smaller molecular compositions.Agar
Sugar is generally heated to 90 DEG C or more in water and dissolves, and temperature forms the gel of good semi-solid when dropping to 35~40 DEG C,
This is the main feature and basis that it serves many purposes.Ago-Gel performance is usually indicated with gel strength.Intensity is higher,
Gelling performance is better.
Agar, agarose especially have significant stability, stagnant degree and hysteresis quality because there is special gelling property, and
Moisture is easily absorbed, there is special stabilizing effect；It is widely used in the fields such as edible, medicine, chemical industry, weaving, national defence, according to not
Statistics completely, agar, the purposes of agarose are more than 1000 kinds existing, can be used for producing in the food industry：Table jelly squares, sizing are soft
Sugar, aquatic products, canned meat, fruit drink, fruit squash, rice wine beverage, dairy beverage, fine work, dairy products cake.Since its is good
Good biocompatibility is widely used in the production of bio-separation medium again.Briefly, the agar pectin in agar is removed, is remained
Under part, be exactly agarose.
At present biomedicine field be commonly used to be separated, the material matrix that purifies can be divided into it is inorganic and organic two
Major class.The medium of inorganic matrix mainly includes silica gel, cellular glass, hydroxyapatite etc., mostly filling out as high performance liquid chromatography
Charge.The medium of organic substrate is usually natural polysaccharide, synthetic high polymer etc., and the matrix that these materials are prepared into is usually pearl ball
Shape.Base ball be various separating mediums it is most basic, most important " raw material ".In bioengineering, medicine preparation or even electronic information
The fields such as industry all have been widely used.Currently, the preparation method of Agarose Magnetic Microsphere has limitation very much, mainly
Using the methods of ultrasound, mechanical stirring, homogenizer emulsification.It is seen that there are no a kind of fine jades of method preparation in existing method
Lipolysaccharide magnetic microsphere can achieve the requirement of perfect microspheres, limit Agarose Magnetic Microsphere in medical treatment, bio-separation purifying
Extensive use.Inverse suspension method is usually that small magnetic-particle is embedded in fine jade by the inverse suspension method of Water-In-Oil at present
Magnetic Agarose sugar microballoon is obtained in lipolysaccharide microballoon, although Magnetic Agarose sugar microballoon made from this method is magnetic preferably, partial size compared with
Greatly, typically larger than 20 μm, and particle diameter distribution is very wide.Therefore, a kind of agar polysaccharide magnetic microsphere is invented to separation of biopolymer
Technical field of purification has positive effect.
Summary of the invention
Present invention mainly solves the technical issues of, for current inverse suspension method be usually by small magnetic-particle, pass through
The inverse suspension method of Water-In-Oil is embedded in agarose microbeads and obtains Magnetic Agarose sugar microballoon, Magnetic Agarose made from this method
Although sugared microballoon is magnetic, preferably partial size is larger, typically larger than 20 μm, and the defect that particle diameter distribution is very wide, provide a kind of fine jade
The preparation method of lipopolysaccharides magnetic microsphere.
In order to solve the above-mentioned technical problem, the technical scheme adopted by the invention is that：
A kind of preparation method of agar polysaccharide magnetic microsphere, it is characterised in that specifically preparation step is：
（1）It is three be packed into after 30% hydrogenperoxide steam generator mixes with blender and thermometer by frerrous chloride and mass fraction
In mouth flask, starting blender is stirred to react, and obtains pre-reaction liquid；
（2）Reaction substrate is obtained after the methanol solution that above-mentioned pre-reaction liquid and mass fraction are 80% is mixed, reaction substrate is filled
Enter bottom to have in the autoclave of aerator, then aluminum chloride powder is added into reaction kettle, seals reaction kettle, pass through exposure
Device of air is passed through ammonia into reaction substrate, until reactor pressure reaches 2.4~2.5MPa；
（3）The temperature for increasing aforesaid reaction vessel is 420~430 DEG C, 1~2h of high-temperature high-voltage reaction, after reaction, reduces reaction
Kettle temperature degree is stirred to react 30~40min, obtains pre-product to 70~80 DEG C；
（4）Polysaccharide solution is obtained after agar polysaccharide and distilled water are mixed, above-mentioned pre-product and polysaccharide solution are mixed to get water
Phase, then toluene, chloroform and Si Pan 80 are mixed to prepare oily phase；
（5）Above-mentioned water phase after being stirred to react, is separated with oil mutually by isometric than being fitted into high-speed mixer after mixing with magnetic field
It is precipitated, is successively rinsed after 3~5 times respectively with ether and water and be drying to obtain agar polysaccharide magnetic microsphere.
Step（1）Described in frerrous chloride and mass fraction be the mass ratio of 30% hydrogenperoxide steam generator be 1:5, stirring
The revolving speed of reaction is 200~300r/min, and the time being stirred to react is 15~20min.
Step（2）Described in pre-reaction liquid and mass fraction be 80% the mass ratio of methanol solution be 2:1, aluminium chloride
The additional amount of powder is the 1% of reaction substrate quality.
Step（3）Described in the revolving speed being stirred to react be 300~400r/min.
Step（4）Described in agar polysaccharide and distilled water mass ratio be 1:40, the quality of pre-product and polysaccharide solution
Than being 1:30, the volume ratio of toluene, chloroform and Si Pan 80 is 20:10:1.
Step（5）Described in the temperature being stirred to react be 60~70 DEG C, the revolving speed being stirred to react be 2000~3000r/
Min, the time being stirred to react are 10~15min.
The beneficial effects of the invention are as follows：
The present invention by frerrous chloride and hydroperoxidation, obtains pre-reaction liquid first, and pre-reaction liquid and methanol are mixed to get
Reaction substrate, then ammonia is passed through into reaction substrate, reaction obtains pre-product under conditions of aluminium oxide and high temperature and pressure, will be pre-
Product and agar polysaccharide solution are mixed to get liquid phase, and toluene, chloroform are mixed to get oily phase, finally by inverse suspension legal system
Agar polysaccharide magnetic microsphere, the present invention prepares magnetic iron oxide powder by raw material of frerrous chloride, during preparation, to pre-
It joined methanol in reaction solution, using methanol as raw material, high-temperature high-voltage reaction carried out under the action of alumina catalyst, and be passed through
Ammonia, so that methanol and ammonia reaction generate organic amine, and the polar group being rich in organic amine has certain electron supplying capacity,
Coordinate bond can be formed between iron ion and chemisorption occurs, thus iron ion and ferrous ion surface in pre-reaction liquid
One layer of monomolecular film is formed, this layer of monomolecular film can provide steric hindrance, prevent magnetic iron oxide particles partial size excessive, simultaneously
The ammonia that the present invention is passed through, which is dissolved in after water, to be obtained ammonium hydroxide and can be used as coprecipitator, and finally obtained partial size is small, and size uniformity is received
Meter level magnetic microsphere is wrapped in nano-scale magnetic by hydrogen bond, coordinate bond absorption using agarose finally by inverse phase suspension
On microballoon crystal grain, close chondritic is formed, is finally made agar polysaccharide magnetic microsphere, agar polysaccharide magnetic produced by the present invention
Property microspherulite diameter is small, is evenly distributed, has broad application prospects.
Specific embodiment
It is in mass ratio 1:Frerrous chloride and mass fraction are had stirring to be packed into after the mixing of 30% hydrogenperoxide steam generator by 5
In the three-necked flask of device and thermometer, starts blender with the revolving speed of 200~300r/min and be stirred to react 15~20min, obtain
Pre-reaction liquid；It is in mass ratio 2 by the methanol solution that above-mentioned pre-reaction liquid and mass fraction are 80%:Reaction bottom is obtained after 1 mixing
Reaction substrate is fitted into bottom and had in the autoclave of aerator, then reaction substrate quality is added into reaction kettle by object
1% aluminum chloride powder seals reaction kettle, is passed through ammonia into reaction substrate by aerator, until reactor pressure reaches
2.4~2.5MPa；The temperature for increasing aforesaid reaction vessel is 420~430 DEG C, 1~2h of high-temperature high-voltage reaction, after reaction, drop
Low reaction kettle temperature degree is stirred to react 30~40min to 70~80 DEG C, with the revolving speed of 300~400r/min, obtains pre-product；By fine jade
Lipopolysaccharides and distilled water are 1 in mass ratio:Polysaccharide solution is obtained after 40 mixing, is in mass ratio 1:30 by above-mentioned pre-product and more
Sugar juice is mixed to get water phase, then by toluene, chloroform and Si Pan 80 is by volume 20:10:1 is mixed to prepare oily phase；It will
Above-mentioned water phase is mutually pressed with oil in equal volume than being fitted into high-speed mixer after mixing, with 2000~3000r/min at 60~70 DEG C
Revolving speed be stirred to react 10~15min after, with the isolated precipitating in magnetic field, done after successively rinsing 3~5 times respectively with ether and water
Dry agar polysaccharide magnetic microsphere to obtain the final product.
It is in mass ratio 1:Frerrous chloride and mass fraction are had stirring to be packed into after the mixing of 30% hydrogenperoxide steam generator by 5
In the three-necked flask of device and thermometer, starts blender with the revolving speed of 200r/min and be stirred to react 15min, obtain pre-reaction liquid；
It is in mass ratio 2 by the methanol solution that above-mentioned pre-reaction liquid and mass fraction are 80%:Reaction substrate is obtained after 1 mixing, will be reacted
Substrate is fitted into bottom and has in the autoclave of aerator, then the aluminium chloride of reaction substrate quality 1% is added into reaction kettle
Powder seals reaction kettle, is passed through ammonia into reaction substrate by aerator, until reactor pressure reaches 2.4MPa；It rises
The temperature of high aforesaid reaction vessel be 420 DEG C, high-temperature high-voltage reaction 1h, after reaction, reduce temperature of reaction kettle to 70 DEG C, with
The revolving speed of 300r/min is stirred to react 30min, obtains pre-product；It is in mass ratio 1 by agar polysaccharide and distilled water:After 40 mixing
Polysaccharide solution is obtained, is in mass ratio 1:Above-mentioned pre-product and polysaccharide solution are mixed to get water phase by 30, then by toluene, three chloromethanes
Alkane and Si Pan 80 are 20 by volume:10:1 is mixed to prepare oily phase；By above-mentioned water phase and oil mutually by isometric than being packed into after mixing
In high-speed mixer, after the revolving speed at 60 DEG C with 2000r/min is stirred to react 10min, with the isolated precipitating in magnetic field, successively
It is rinsed after 3 times respectively with ether and water and is drying to obtain agar polysaccharide magnetic microsphere.
It is in mass ratio 1:Frerrous chloride and mass fraction are had stirring to be packed into after the mixing of 30% hydrogenperoxide steam generator by 5
In the three-necked flask of device and thermometer, starts blender with the revolving speed of 250r/min and be stirred to react 17min, obtain pre-reaction liquid；
It is in mass ratio 2 by the methanol solution that above-mentioned pre-reaction liquid and mass fraction are 80%:Reaction substrate is obtained after 1 mixing, will be reacted
Substrate is fitted into bottom and has in the autoclave of aerator, then the aluminium chloride of reaction substrate quality 1% is added into reaction kettle
Powder seals reaction kettle, is passed through ammonia into reaction substrate by aerator, until reactor pressure reaches 2.4MPa；It rises
The temperature of high aforesaid reaction vessel be 425 DEG C, high-temperature high-voltage reaction 1.5h, after reaction, reduce temperature of reaction kettle to 75 DEG C, with
The revolving speed of 350r/min is stirred to react 35min, obtains pre-product；It is in mass ratio 1 by agar polysaccharide and distilled water:After 40 mixing
Polysaccharide solution is obtained, is in mass ratio 1:Above-mentioned pre-product and polysaccharide solution are mixed to get water phase by 30, then by toluene, three chloromethanes
Alkane and Si Pan 80 are 20 by volume:10:1 is mixed to prepare oily phase；By above-mentioned water phase and oil mutually by isometric than being packed into after mixing
In high-speed mixer, after the revolving speed at 65 DEG C with 2500r/min is stirred to react 12min, with the isolated precipitating in magnetic field, successively
It is rinsed after 4 times respectively with ether and water and is drying to obtain agar polysaccharide magnetic microsphere.
It is in mass ratio 1:Frerrous chloride and mass fraction are had stirring to be packed into after the mixing of 30% hydrogenperoxide steam generator by 5
In the three-necked flask of device and thermometer, starts blender with the revolving speed of 300r/min and be stirred to react 20min, obtain pre-reaction liquid；
It is in mass ratio 2 by the methanol solution that above-mentioned pre-reaction liquid and mass fraction are 80%:Reaction substrate is obtained after 1 mixing, will be reacted
Substrate is fitted into bottom and has in the autoclave of aerator, then the aluminium chloride of reaction substrate quality 1% is added into reaction kettle
Powder seals reaction kettle, is passed through ammonia into reaction substrate by aerator, until reactor pressure reaches 2.5MPa；It rises
The temperature of high aforesaid reaction vessel be 430 DEG C, high-temperature high-voltage reaction 2h, after reaction, reduce temperature of reaction kettle to 80 DEG C, with
The revolving speed of 400r/min is stirred to react 40min, obtains pre-product；It is in mass ratio 1 by agar polysaccharide and distilled water:After 40 mixing
Polysaccharide solution is obtained, is in mass ratio 1:Above-mentioned pre-product and polysaccharide solution are mixed to get water phase by 30, then by toluene, three chloromethanes
Alkane and Si Pan 80 are 20 by volume:10:1 is mixed to prepare oily phase；By above-mentioned water phase and oil mutually by isometric than being packed into after mixing
In high-speed mixer, after the revolving speed at 70 DEG C with 3000r/min is stirred to react 15min, with the isolated precipitating in magnetic field, successively
It is rinsed after 5 times respectively with ether and water and is drying to obtain agar polysaccharide magnetic microsphere.
The agar polysaccharide magnetic microsphere that comparative example is produced with Gansu company is as a comparison case to agar produced by the present invention
Agar polysaccharide magnetic microsphere in polysaccharide magnetic microsphere and comparative example carries out performance detection, and testing result is as shown in table 1：
Test method：
Particle size test is detected using laser particle size analyzer.
Magnetic saturation intensity test is used to be detected than magnetic saturation intensity analyzer.
1 agar polysaccharide magnetic microsphere performance measurement result of table
| Test item | Example 1 | Example 2 | Example 3 | Comparative example |
| Partial size（μm） | 4 | 3 | 2 | 18 |
| Magnetic saturation intensity（emu/g） | 21 | 21 | 22 | 10 |
According to data among the above, agar polysaccharide magnetic microsphere magnetism of the invention is good, and uniform particle diameter, particle diameter distribution is narrow, with city
It is smaller compared to its partial size to sell product, at 2~4 μm, specific surface area is bigger, and active site is more, has broad application prospects.
Claims (6)
1. a kind of preparation method of agar polysaccharide magnetic microsphere, it is characterised in that specifically preparation step is：
（1）It is three be packed into after 30% hydrogenperoxide steam generator mixes with blender and thermometer by frerrous chloride and mass fraction
In mouth flask, starting blender is stirred to react, and obtains pre-reaction liquid；
（2）Reaction substrate is obtained after the methanol solution that above-mentioned pre-reaction liquid and mass fraction are 80% is mixed, reaction substrate is filled
Enter bottom to have in the autoclave of aerator, then aluminum chloride powder is added into reaction kettle, seals reaction kettle, pass through exposure
Device of air is passed through ammonia into reaction substrate, until reactor pressure reaches 2.4~2.5MPa；
（3）The temperature for increasing aforesaid reaction vessel is 420~430 DEG C, 1~2h of high-temperature high-voltage reaction, after reaction, reduces reaction
Kettle temperature degree is stirred to react 30~40min, obtains pre-product to 70~80 DEG C；
（4）Polysaccharide solution is obtained after agar polysaccharide and distilled water are mixed, above-mentioned pre-product and polysaccharide solution are mixed to get water
Phase, then toluene, chloroform and Si Pan 80 are mixed to prepare oily phase；
（5）Above-mentioned water phase after being stirred to react, is separated with oil mutually by isometric than being fitted into high-speed mixer after mixing with magnetic field
It is precipitated, is successively rinsed after 3~5 times respectively with ether and water and be drying to obtain agar polysaccharide magnetic microsphere.
2. a kind of preparation method of agar polysaccharide magnetic microsphere according to claim 1, it is characterised in that：Step（1）In
The frerrous chloride and mass fraction is that the mass ratio of 30% hydrogenperoxide steam generator is 1:5, the revolving speed being stirred to react be 200~
300r/min, the time being stirred to react are 15~20min.
3. a kind of preparation method of agar polysaccharide magnetic microsphere according to claim 1, it is characterised in that：Step（2）In
The mass ratio for the methanol solution that the pre-reaction liquid and mass fraction are 80% is 2:1, the additional amount of aluminum chloride powder is reaction
The 1% of substrate quality.
4. a kind of preparation method of agar polysaccharide magnetic microsphere according to claim 1, it is characterised in that：Step（3）In
The revolving speed being stirred to react is 300~400r/min.
5. a kind of preparation method of agar polysaccharide magnetic microsphere according to claim 1, it is characterised in that：Step（4）In
The mass ratio of the agar polysaccharide and distilled water is 1:40, the mass ratio of pre-product and polysaccharide solution is 1:30, toluene, trichlorine
The volume ratio of methane and Si Pan 80 are 20:10:1.
6. a kind of preparation method of agar polysaccharide magnetic microsphere according to claim 1, it is characterised in that：Step（5）In
The temperature being stirred to react is 60~70 DEG C, and the revolving speed being stirred to react is 2000~3000r/min, the time being stirred to react
For 10~15min.
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Citations (5)
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|---|---|---|---|---|
| CN1376730A (en) * | 2001-03-27 | 2002-10-30 | 内蒙古师范大学 | Process for preparing magnetic composite agarose microspheres |
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| CN102994489A (en) * | 2012-10-09 | 2013-03-27 | 山西农业大学 | Biotin-avidin system immobilized glucoamylase and preparation method thereof |
| CN103483602A (en) * | 2013-09-16 | 2014-01-01 | 海狸纳米科技（苏州）有限公司 | Preparation method for small-particle-size magnetic agarose microspheres |
| CN103007847B (en) * | 2012-12-20 | 2014-12-31 | 华南理工大学 | Magnetic nanoparticle-based immobilized laccase and ionic liquid composite particle and application thereof |
-
2018
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|---|---|---|---|---|
| CN1376730A (en) * | 2001-03-27 | 2002-10-30 | 内蒙古师范大学 | Process for preparing magnetic composite agarose microspheres |
| CN1979699A (en) * | 2006-11-09 | 2007-06-13 | 上海交通大学 | Method for preparing iron-oxide particle suspension |
| CN102994489A (en) * | 2012-10-09 | 2013-03-27 | 山西农业大学 | Biotin-avidin system immobilized glucoamylase and preparation method thereof |
| CN103007847B (en) * | 2012-12-20 | 2014-12-31 | 华南理工大学 | Magnetic nanoparticle-based immobilized laccase and ionic liquid composite particle and application thereof |
| CN103483602A (en) * | 2013-09-16 | 2014-01-01 | 海狸纳米科技（苏州）有限公司 | Preparation method for small-particle-size magnetic agarose microspheres |
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