CN105968825A - Method for preparing cellular material from biological cross-linking agent crosslinked protein - Google Patents
Method for preparing cellular material from biological cross-linking agent crosslinked protein Download PDFInfo
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- CN105968825A CN105968825A CN201610375163.7A CN201610375163A CN105968825A CN 105968825 A CN105968825 A CN 105968825A CN 201610375163 A CN201610375163 A CN 201610375163A CN 105968825 A CN105968825 A CN 105968825A
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
- C08L89/00—Compositions of proteins; Compositions of derivatives thereof
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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
- C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
- C08J3/246—Intercrosslinking of at least two polymers
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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
- C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
- C08J9/04—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof using blowing gases generated by a previously added blowing agent
- C08J9/12—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof using blowing gases generated by a previously added blowing agent by a physical blowing agent
- C08J9/14—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof using blowing gases generated by a previously added blowing agent by a physical blowing agent organic
- C08J9/141—Hydrocarbons
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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
- C08J2203/00—Foams characterized by the expanding agent
- C08J2203/14—Saturated hydrocarbons, e.g. butane; Unspecified hydrocarbons
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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
- C08J2389/00—Characterised by the use of proteins; Derivatives thereof
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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
- C08J2405/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2401/00 or C08J2403/00
- C08J2405/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
- C08L2201/00—Properties
- C08L2201/06—Biodegradable
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- Chemical Kinetics & Catalysis (AREA)
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- Polymers & Plastics (AREA)
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Abstract
The invention discloses a method for preparing a cellular material from biological cross-linking agent crosslinked protein. The method comprises the following steps that 1, chitosan is added into an acetic acid solution and is fully hydrated; 2, wheat gliadin is added into an ethanol solution to be fully dissolved; 3, the material obtained in the step 1 and the material obtained in the step 2 are mixed, shear-homogenized, centrifuged and rotationally evaporated through an anti-solvent method, and wheat gliadin compound colloidal particles are obtained; 4, genipin is added into the material obtained in the step 3 to be fully dissolved; 5, normal hexane is added into the material obtained in the step 4 for shearing and homogenizing, and the novel cellular material is prepared after freeze drying. The obtained cellular material is rich in raw material source, low in cost and high in porosity and specific surface area, has certain anti-pressure ability, and has certain prospects in the fields of biological catalysis, chemical industry, medicine, food and the like.
Description
Technical field
The present invention relates to a kind of novel porous materials, particularly related to one and prepared porous with Biological cross-linker crosslinking protein
The method of material.
Technical background
The research of porous material has become as a big study hotspot of Material Field, and porous material is owing to itself having a lot of spy
Different character, such as high porosity, high-specific surface area, high adsorption, high screening property etc., these other materials do not have
Character makes porous material have a wide range of applications at numerous areas such as chemical industry, functional material, medical science, environmental protection, food.
The raw material used by porous material developed at present is substantially these things such as metal, pottery, organic-inorganic chemical reagent
Matter, their common feature is all the inedible raw material used, or has the raw material of certain toxic action to human body.
The cross-linking agent that additionally prior art is used is substantially glutaraldehyde or other chemical cross-linking agents, these cross-linking agent often toxicity
Very big, be not suitable for the application in a lot of field.
The method not finding in prior art to utilize genipin crosslinking protein and chitosan to prepare porous material.Existing document
(Nadeem Siddiqui,Esmaiel Jabbari,et al.Osteogenic differentiation of human mesenchymal
stem cells in freeze-gelled chitosan/nanoβ-tricalcium phosphate porous scaffolds crosslinked
With genipin [J] .Materials Science and Engineering:C.2015,54:79-83) utilize bata-tricalcium phosphate and shell to gather
Sugar is raw material, and genipin prepares porous material for human bone timbering material as cross-linking agent, and it is only used for medical science bone support
Material and the fields such as biological food cannot be applied to, be of limited application.
Summary of the invention
It is an object of the invention to overcome the shortcoming of prior art, it is provided that one need not use toxic cross-linking agent, use human body
The raw material of nonhazardous effect, the method preparing porous material with Biological cross-linker crosslinking protein, raw material used in the present invention is equal
For edible, having good biocompatibility, the porosity of gained porous material is more than 90%, has the anti-of more than 20kPa
Compressive Strength.
Genipin is jasminoidin product after β glucoside enzyme hydrolysis, is a kind of excellent natural biological cross-linking agent, permissible
Cross-linking with albumen, gelatin and chitosan etc., the raw material that the present invention uses is wheat gliadin and chitosan, positive good utilisation capital
It is feature crosslinking protein and the chitosan of splendid natural biological cross-linking agent that Buddhist nun puts down, and makes porous material have certain mechanical property.
The object of the invention is realized by following technical scheme.
A kind of method preparing porous material with Biological cross-linker crosslinking protein, comprises the following steps:
(1) prepare the acetum that acetic acid quality concentration is 0.1%-5%, will be equivalent to acetum quality 0.01%-3%
Chitosan adds in acetum so that it is fully aquation.
(2) configuration ethanol mass concentration is the ethanol solution of 30%-90%, will be equivalent to ethanol solution quality 0.5%-5%
Wheat gliadin adds in ethanol solution makes it fully dissolve.
(3) material of step (2) gained taking the 20%-200% being equivalent to the volume of material that step (1) obtains adds step
Suddenly in the material of (1), 3000r/min-30000r/min rotating speed down cut homogenizing.
(4) by the material of step (3) gained by water-bath rotary evaporation to the mass concentration of wheat gliadin composite particles
For 0.1%-5%.
(5) material of step (4) gained is centrifuged under 2000g-10000g rotating speed, abandons precipitation and take supernatant.
(6) adding the genipin that quality is albumen 1%-10% in step (5) gained material, stirring makes it fully dissolve.
(7) take the normal hexane that volume of material accounting is 30%-90% obtained with step (6) and add what step (6) obtained
In material, under 3000r/min-30000r/min rotating speed, fully shear homogenizing 1-10min.
(8) the material lyophilization of step (7) gained is i.e. prepared a kind of porous material with Biological cross-linker crosslinking protein
Material.
Porous material prepared by the present invention is because having the porosity of more than 90% and the comprcssive strength of more than 20kPa, and material has simultaneously
Have the feature of edibility, be expected to be applied to as carrier high-efficiency adsorption activity material or medicine food, biology and targeting to
Medicine medical field, applied range.
Of the present invention a kind of prepare the method for porous material with Biological cross-linker crosslinking protein and have the following advantages and useful effect
Really:
(1) the raw material albumen source employed in the preparation method of wheat gliadin porous material of the present invention is natural,
Without any side effects to human body.Prepare the former material that the chitosan used by wheat gliadin composite nanometer particle is also edibility
Material, the most basic for dissolving the normal hexane of chitosan, the acetum of wheat gliadin and ethanol solution and follow-up use
Volatilization completely, seldom remains, to human body without any harm.
(2) use the natural porous material prepared by the crosslinking of Biological cross-linker genipin that this porous material is had necessarily
Mechanical property, simultaneously compared to glutaraldehyde or other chemical cross-linking agents, genipin source is natural without any side effects.
(3) commercially available porous material can produce a large amount of pollutant in process of manufacture, and porous material discarded after using
Material is difficult to degrade at nature, long-term pollution environment.Along with development and people's raising to environmental requirement of science and technology, pass
The porous material of system is increasingly difficult in adapt to the needs of current economic development and daily life, and one of the present invention is handed over biology
Connection agent crosslinking protein prepare porous material prepared in the method for porous material due to raw material sources natural to environment without any
Pollute, do not deposit the problem being difficult to after producing pollution in process of production and discarding degrade.
(4) present invention is a kind of novel porous materials from the exploitation of the bond material field, visual angle of food, has both reached food
Standard has again the performance of material, and its preparation technology is simple, and raw material is cheap, will play a significant role at numerous areas.
Accompanying drawing explanation
Fig. 1 is the scanning electron of porous material prepared by 2% wheat gliadin colloidal solid and 70% normal hexane in embodiment 1
Microscope figure；
Fig. 2 is Fig. 1 scanning electron microscope partial enlarged drawing.
Detailed description of the invention
For being more fully understood that the present invention, the present invention will be further described with embodiment below in conjunction with the accompanying drawings, but the guarantor of the present invention
The scope of protecting is not limited to the scope of embodiment statement.
The porosity of the porous material in the embodiment of the present invention is full-automatic high by Merck & Co., Inc of U.S. Auto Pore IV 9500
Performance mercury injection apparatus is measured, and the mechanical property of material is tested by AG-IC50kN electronic universal tester.Embodiment is medium and small
The mass concentration of gliadin composite colloid granule is constantly to be claimed by electronic balance during rotary evaporation when preparing granule
Measure the change of its quality to control the mass concentration of granule.
Embodiment 1
A kind of method preparing porous material with Biological cross-linker crosslinking protein, comprises the following steps:
(1) preparing the acetum that acetic acid quality concentration is 1%, the chitosan that will be equivalent to acetum quality 0.05% adds
Enter in acetum so that it is fully aquation.
(2) configuration ethanol mass concentration is the ethanol solution of 70%, and the Semen Tritici aestivi alcohol that will be equivalent to ethanol solution quality 2.5% is molten
Albumen adds in ethanol solution makes it fully dissolve.
(3) material of step (2) gained taking be equivalent to the volume of material that step (1) obtains 40% adds step (1)
Material in, 6000r/min rotating speed down cut homogenizing.
(4) by the material of step (3) gained by water-bath rotary evaporation to the mass concentration of wheat gliadin composite particles
It is 2%.
(5) material of step (4) gained is centrifuged under 8000g rotating speed, abandons precipitation and take supernatant.
(6) adding the genipin that quality is albumen 5% in step (5) gained material, stirring makes it fully dissolve.
(7) take the normal hexane that volume of material accounting is 70% obtained with step (6) and add in the material that step (6) obtains,
Homogenizing 1min is fully sheared under 10000r/min rotating speed.
(8) by the material lyophilization of step (7) gained, a kind of porous with Biological cross-linker crosslinking protein is i.e. prepared
Material.
The mass concentration of wheat gliadin composite particles in step (4) has been carried out gradient test, and mass concentration is respectively as follows:
0.1%, 0.5%, 1.0%, 2.0%, 3.0%, wherein the granule density of 0.1% fails to form High Internal Phase Emulsion thus cannot make
Standby porous material, porous material prepared by the present embodiment measures its porosity by mercury injection apparatus, test result such as table 1.
Table 1
Owing to the difference of granule density directly affects the oil-water interfaces structure of granule and the formed emulsion of normal hexane, thus affect
The character of the porous material after lyophilization, the porous material that as can be seen from Table 1 prepared by the present embodiment has the highest hole
Gap rate, wherein the porosity of 1.0% granule density is the highest, it may be possible to be the porous material hole knot formed due to granule density height
The more abundant fraction porosity of structure is relatively low, and when granule density is relatively low, the pore structure of material relatively lean aperture larger porosity is lower.
In the present embodiment, the mass concentration of wheat gliadin composite colloid granule is the most open close during rotary evaporation when preparing granule
Cross what the change of its quality of electronic balance weighing controlled.
The present embodiment uses genipin crosslinking, prepared porous material, has the anti-of 40kPa through universal testing machine test
Compressive Strength, this comprcssive strength makes the loose structure of material energy holding itself be unlikely to subside, be preferably applied for food,
Biology and target administration medical field.
The scanning electron microscope of the porous material that Fig. 1 is 2% wheat gliadin colloidal solid and 70% normal hexane is prepared
Figure, as can be seen from the figure prepared material is dispersed with the hole of densification, has the highest porosity.Fig. 2 is its office
Portion's enlarged drawing, it can be seen that the pore structure of this porous material presents round and smooth abundant, this has with the compressive property of porous material
Certain relation.
The raw material (wheat gliadin, chitosan, genipin) used by porous material that the present embodiment prepares simultaneously
Be edibility, for dissolve chitosan, the acetum of wheat gliadin and ethanol solution and follow-up use just oneself
Alkane volatilizees completely the most substantially, seldom remains, and to human body without any harm, fullys meet edible state.
Prior art (C.L.Zhou, Ngai, et al.Fabrication and characterization of pure porous Ti3SiC2
With controlledporosity and pore features [J] .Materials Letters, 2014,131:280-283.) porous prepared
Material not only porosity relatively low (75%), and be basically applied to the Material Fields such as ceramic metal and food, life cannot be applied to
Thing and medical system field.Porous material prepared by the present invention has the highest porosity (more than 90%) and edibility,
It is expected to be applied to food, biology and target administration medical field as carrier high-efficiency adsorption activity material or medicine.
Embodiment 2
A kind of method preparing porous material with Biological cross-linker crosslinking protein, comprises the following steps:
(1) preparing the acetum that acetic acid quality concentration is 1%, the chitosan that will be equivalent to acetum quality 0.05% adds
Enter in acetum so that it is fully aquation.
(2) configuration ethanol mass concentration is the ethanol solution of 70%, and the Semen Tritici aestivi alcohol that will be equivalent to ethanol solution quality 2.5% is molten
Albumen adds in ethanol solution makes it fully dissolve.
(3) material of step (2) gained taking be equivalent to the volume of material that step (1) obtains 40% adds step (1)
Material in, 6000r/min rotating speed down cut homogenizing.
(4) by the material of step (3) gained by water-bath rotary evaporation to the mass concentration of wheat gliadin composite particles
It is 2%.
(5) material of step (4) gained is centrifuged under 8000g rotating speed, abandons precipitation and take supernatant.
(6) adding the genipin that quality is albumen 2% in step (5) gained material, stirring makes it fully dissolve.
(7) take the normal hexane that volume of material accounting is 70% obtained with step (6) and add in the material that step (6) obtains,
Homogenizing 1min is fully sheared under 10000r/min rotating speed.
(8) the material lyophilization of step (7) gained is i.e. prepared a kind of porous material with Biological cross-linker crosslinking protein
Material.
Porosity of porous material prepared by the present embodiment is up to 92.3216%, and has the comprcssive strength of 20kPa, the most right
In step (6), wheat gliadin quality shared by genipin has carried out gradient test, proportion is respectively as follows: 0,1%, 2%,
5%, 10%.Corresponding porosity test result such as table 2, it can be seen that the porosity of porous material of 2% genipin crosslinking is the highest,
When being possibly due to crosslinker concentration height, pore forming process is had certain resistance and to show porosity relatively low.
Table 2
Embodiment 3
A kind of method preparing porous material with Biological cross-linker crosslinking protein, comprises the following steps:
(1) preparing the acetum that acetic acid quality concentration is 1%, the chitosan that will be equivalent to acetum quality 0.05% adds
Enter in acetum so that it is fully aquation.
(2) configuration ethanol mass concentration is the ethanol solution of 70%, and the Semen Tritici aestivi alcohol that will be equivalent to ethanol solution quality 2.5% is molten
Albumen adds in ethanol solution makes it fully dissolve.
(3) material of step (2) gained taking be equivalent to the volume of material that step (1) obtains 40% adds step (1)
Material in, 6000r/min rotating speed down cut homogenizing.
(4) by the material of step (3) gained by water-bath rotary evaporation to the mass concentration of wheat gliadin composite particles
It is 2%.
(5) material of step (4) gained is centrifuged under 8000g rotating speed, abandons precipitation and take supernatant.
(6) adding the genipin that quality is albumen 5% in step (5) gained material, stirring makes it fully dissolve.
(7) take the normal hexane that volume of material accounting is 90% obtained with step (6) and add in the material that step (6) obtains,
Homogenizing 1min is fully sheared under 10000r/min rotating speed.
(8) the material lyophilization of step (7) gained is i.e. prepared a kind of porous material with Biological cross-linker crosslinking protein
Material.
Porosity of porous material prepared by the present embodiment is up to 97.4612%, and has the comprcssive strength of 25kPa, the most right
Volume ratio shared by step (7) normal hexane has carried out gradient test, proportion is respectively as follows: 50%, 60%, 70%, 80%,
90%.Corresponding porosity test result such as table 3, due to the emulsion formed when normal hexane volume accounting is by 50% and 60%
Oil phase is the lowest and is layered, it is impossible to prepare porous material；Can be seen that the porosity of porous material of the highest preparation of oil phase simultaneously
The highest, this is that the internal phase removed when preparing porous material is the most owing to oil phase accounting is the biggest, and the material porosity of formation is more
High.
Table 3
The foregoing is only embodiments of the invention, not thereby limit the scope of the claims of the present invention, every present invention of utilization says
Equivalent structure that bright book content is done or equivalence flow process exchange, or be directly or indirectly used in other association area, all with
Reason is included in the scope of patent protection of the present invention.
Claims (9)
1. the method preparing porous material with Biological cross-linker crosslinking protein, it is characterised in that comprise the steps:
(1) configuration acetum, the chitosan that will be equivalent to acetum quality 0.01%-3% adds in acetum, fills
Divide aquation；
(2) configuration ethanol solution, the wheat gliadin that will be equivalent to ethanol solution quality 0.5%-5% adds in ethanol solution,
Stirring, fully dissolves；
(3) step (2) gained of the 20%-200% being equivalent to the volume of material that step (1) obtains is taken by anti-solvent method
Material, add step (1) material in, shear homogenizing；
(4) by the material of step (3) gained by the mass concentration being evaporated to wheat gliadin composite colloid granule it is
0.1%-5%；
(5) material of step (4) gained is centrifuged, abandons precipitation and take supernatant；
(6) genipin is added in step (5) gained material, fully dissolve；
(7) normal hexane is added in the material that step (6) obtains, fully shear homogenizing；
(8) by the material lyophilization of step (7) gained, the porous material of Biological cross-linker crosslinking protein is obtained.
The method that Biological cross-linker crosslinking protein the most according to claim 1 prepares porous material, it is characterised in that institute
Stating acetic acid quality concentration in step (1) is 0.1%-5%.
The method that Biological cross-linker crosslinking protein the most according to claim 1 prepares porous material, it is characterised in that institute
Stating ethanol mass concentration in step (2) is 30%-90%.
The method that Biological cross-linker crosslinking protein the most according to claim 1 prepares porous material, it is characterised in that step
Suddenly (3) and the rotating speed shearing homogenizing described in step (7) are all 3000r/min 30000r/min.
The method that Biological cross-linker crosslinking protein the most according to claim 1 prepares porous material, it is characterised in that institute
State and be evaporated to water-bath rotary evaporation.
The method that Biological cross-linker crosslinking protein the most according to claim 5 prepares porous material, it is characterised in that institute
The temperature stating water-bath rotary evaporation is 25 DEG C-80 DEG C, and rotating speed is 50r/min 200r/min.
The method that Biological cross-linker crosslinking protein the most according to claim 1 prepares porous material, it is characterised in that institute
Stating centrifugal rotating speed is 2000g-10000g.
The method that Biological cross-linker crosslinking protein the most according to claim 1 prepares porous material, it is characterised in that step
Suddenly the 1%-10% of wheat gliadin quality during the consumption of genipin described in (6) is step (2).
The method that Biological cross-linker crosslinking protein the most according to claim 1 prepares porous material, it is characterised in that step
Suddenly volume ratio 50%-90% shared by the consumption of normal hexane described in (7).
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| CN106750575A (en) * | 2016-11-29 | 2017-05-31 | 武汉大学 | Hydroxypropyl chitosan/soybean protein isolate composite membrane and preparation method and application |
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| CN115715588A (en) * | 2022-07-27 | 2023-02-28 | 中国农业大学 | Preparation method and application of hordein-chitosan high internal phase Pickering emulsion |
| CN117159729A (en) * | 2023-09-18 | 2023-12-05 | 大连医科大学附属第一医院 | Porous stem cell-loaded biological material and application thereof in pain treatment |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106750575A (en) * | 2016-11-29 | 2017-05-31 | 武汉大学 | Hydroxypropyl chitosan/soybean protein isolate composite membrane and preparation method and application |
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| CN115715588A (en) * | 2022-07-27 | 2023-02-28 | 中国农业大学 | Preparation method and application of hordein-chitosan high internal phase Pickering emulsion |
| CN117159729A (en) * | 2023-09-18 | 2023-12-05 | 大连医科大学附属第一医院 | Porous stem cell-loaded biological material and application thereof in pain treatment |
| CN117159729B (en) * | 2023-09-18 | 2024-04-02 | 大连医科大学附属第一医院 | Porous stem cell-loaded biological material and application thereof in pain treatment |
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