CN107937384A - A kind of preparation method of pickering emulsion stablized based on hydrophobically modified magnetic particle and its immobilised enzymes application - Google Patents
A kind of preparation method of pickering emulsion stablized based on hydrophobically modified magnetic particle and its immobilised enzymes application Download PDFInfo
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Abstract
Preparation method and its immobilised enzymes application the present invention relates to a kind of water-in-oil type pickering emulsion stablized based on hydrophobically modified magnetic particle.With magnetic Fe made from a step hydrophobically modified method3O4@SiO2Particle used as stabilizers, n-dodecane or normal heptane make oil phase, and Tris buffer solutions make water phase, stir above-mentioned mixed liquor using high-shear homogenizer, Fe is made3O4@SiO2The water-in-oil type pickering emulsion that nano-particle is stablized.The pickering emulsion drop stability stablized in the present invention by hydrophobically modified magnetic particle is good, it can be achieved that the cladding of a variety of enzymes, and immobilised enzymes has good heat endurance, storage stability.The pickering emulsion that four kinds of hydrophobically modified magnetic particles are stablized can realize magnetic recovery, significantly improve immobilised enzymes enzyme activity and reusing.Pickering emulsion cost of material of the invention is low, preparation method is easily operated, there is good economic feasibility and practicality.
Description
Technical field
The present invention relates to a kind of preparation method for the water-in-oil type pickering emulsion stablized based on hydrophobically modified magnetic particle
And its immobilised enzymes application, belong to emulsifying agent technical field.
Background technology
Pickering emulsion is the lotion stablized by solids, low with cost, small toxicity, be affected by the external environment small etc.
Advantage, can be widely applied in the fields such as enzyme immobilization, regenerative resource and active material cladding.The stabilization of pickering emulsion
Property is determined by many factors such as oil-water ratio, granule content and oil phase types.For the pickering emulsion that solid particle is stablized,
Its stability depends mainly on adsorption capacity of the solid particle on two-phase interface in system, that is, using quiet between particle
Electric repulsion forms one of special steric barrier with stable emulsion.There is emulsifier system in pickering emulsion research at present
It is of high cost, stability of emulsion is insufficient, recycling and reusing it is bad the problems such as.Magnetic Nano material is that one kind has magnetic response
The nano material of performance, it is with good magnetism, light resistance and the absorption to ultraviolet and screen effect.These features all make
It has important application value in catalytic reaction, system separation and recycling.
At present, most widely used in magnetic Nano material is magnetic Fe3O4Particle and its modified particle, these
Particle receives significant attention in biomaterial preparation, resource reclaim and medicine delay control.Because pickering emulsion is in various aspects
The advantage better than conventional emulsion possessed, utilizes Fe3O4The research that nano-particle stablizes pickering emulsion is also gradually carried out, than
Such as：Zhou etc.[1]With Fe3O4Nano-particle used as stabilizers, the solvent of opposed polarity make oil phase and stablize pickering emulsion, the results showed that
Fe3O4Nano-particle can only could form the lotion of stabilization in some nonpolar solvents, and can be only formed the skin of oil-in-water type
Crin lotion.In addition, Zhang etc.[2]Fe is used respectively3O4-SiO2Nanometer rods and Fe3O4@SiO2Nano-particle stablizes pik woods breast
Liquid, it turns out that only Fe3O4-SiO2Nanometer rods can stablize water-in-oil type pickering emulsion, and the Fe of core shell structure3O4@
SiO2Nano-particle can not stablize water-in-oil type pickering emulsion.Its reason is mainly due to this core shell structure Fe3O4@SiO2
The SiO of particle2Clad is than relatively thin, and further modification is not made on surface so that interparticle electrostatic repulsion is too strong, institute
Water-in-oil type pickering emulsion can not be stablized.
Since magnetic oxide particle surface contains substantial amounts of hydroxyl, very strong hydrophily is made it have, is easier to obtain
The pickering emulsion of oil-in-water type is obtained, and has certain difficulty on water-in-oil type pickering emulsion is prepared, but is fixed in enzyme
We, which need to obtain stable water-in-oil type pickering emulsion, in change system is coated on enzyme in water phase, to play certain protection
Effect.Therefore, present invention is generally directed to produced problem in studying at present to Fe3O4The synthetic method and SiO of particle2Cladding
Layer is improved, and utilizes a step hydrophobically modified method[3]With Fe3O4Particle is core coated Si O2, four kinds of magnetic Fes are prepared3O4@
SiO2Particle.Its difference essentially consists in Fe3O4The difference of particle preparation method, i.e., (be denoted as p-Fe including coprecipitation3O4) and it is molten
The hot method of agent (is denoted as s-Fe3O4) two kinds, and SiO2Solvent whether there is the difference of ethanol addition in clad synthesis, i.e., is only made with water
Solvent (is denoted as w-SiO2) and water and alcohol mixture make solvent and (be denoted as e-SiO2), these four magnetic particles are denoted as p- respectively
Fe3O4@w-SiO2、p-Fe3O4@e-SiO2、s-Fe3O4@w-SiO2、s-Fe3O4@e-SiO2.The present invention by magnetic Nano material with
Pickering emulsion combines, and solves the stability deficiency occurred in conventional pickering emulsion research, is unfavorable for recycling, repeats
The problems such as usability is bad.
Bibliography：
[1].Zhou J,Qiao X,Binks B P,et al.Magnetic Pickering emulsions
stabilized by Fe3O4nanoparticles.Langmuir,2011,27(7):3308-3316.
[2].Zhang L,Zhang F,Wang Y-S,et al.Magnetic colloidosomes fabricated
by Fe3O4-SiO2hetero-nanorods.Soft Matter,2011,7(16):7375.
[3].Zhou,W.J.；Fang,L.；Fan,Z.；Albela,B.；Bonneviot,L..Tunable catalysts
for solvent-free biphasic systems:Pickering interfacial catalysts over
amphiphilic silica nanoparticles.J.Am.Chem.Soc.2014,136,4869-4872.
The content of the invention
Present invention aims at develop a kind of preparation method for the pickering emulsion stablized based on hydrophobically modified magnetic particle
And its application study of immobilised enzymes.Invention includes hydrophobically modified magnetic Fe3O4@SiO2The system of particle-stabilised pickering emulsion
Preparation Method, and magnetic Fe3O4@SiO2The application of particle-stabilised pickering emulsion immobilised enzymes.
Concrete technical scheme is as follows：
A kind of preparation method for the water-in-oil type pickering emulsion stablized based on hydrophobically modified magnetic particle, it is characterized in that：
With magnetic Fe made from a step hydrophobically modified method3O4@SiO2Particle[3]Used as stabilizers, n-dodecane or normal heptane make oil phase, Tris
Buffer solution makees water phase, stirs above-mentioned mixed liquor using high-shear homogenizer, Fe is made3O4@SiO2The water-in-oil type that nano-particle is stablized
Pickering emulsion.
Wherein, magnetic Fe3O4@SiO2The volume fraction that particle accounts for water phase is 0.5-1.5% (%, w/v), water-oil phase body
Product is than being 2:1-20:1, high-shear homogenizer mixing speed is 10000-20000rpm.
The magnetic Fe3O4@SiO2Particle includes：p-Fe3O4@w-SiO2、p-Fe3O4@e-SiO2、s-Fe3O4@w-
SiO2、s-Fe3O4@e-SiO2Four kinds of particles.
The magnetic pickering emulsion is s-Fe3O4@e-SiO2Particle-stabilised pickering emulsion.
Tris buffer solutions are 10mM, pH 8.5.
The water-oil phase volume ratio of the magnetic pickering emulsion is 10:1, magnetic Fe3O4@SiO2Particle accounts for water phase
Volume fraction is 1% (%, w/v), and high-shear homogenizer mixing speed is 20000rpm.
Hydrophobically modified magnetic Fe prepared by the present invention3O4@SiO2Particle-stabilised pickering emulsion answering on immobilised enzymes
With：It is derived from and adds the dilution of Tris buffer solutions by enzyme solutions, makees water phase, magnetic Fe with the enzyme solutions after dilution3O4@SiO2Particle is made
Stabilizer, n-dodecane make oil phase, and using the above-mentioned mixed liquor of homogenizer high-speed stirred, enzyme is coated on water phase in preparation process
In, obtain the magnetic pickering emulsion containing immobilised enzymes；Wherein, free enzyme solutions addition is 10-50 μ L, magnetic Fe3O4@
SiO2The volume fraction that particle accounts for water phase is 0.5-1.5% (%, w/v), and water-oil phase volume ratio is 2:1-20:1, high speed homogenization
Machine mixing speed is 10000-20000rpm.
The free enzyme solutions are liquid aliphatic enzyme.
The present invention relates to a kind of preparation method for the water-in-oil type pickering emulsion stablized based on hydrophobically modified magnetic particle
And its immobilised enzymes application.Two kinds of Fe of coprecipitation and solvent structure are used in the invention respectively3O4Nano-particle, Zhi Houyou
Respectively the magnetic Fe of four kinds of different structures has been obtained as solvent with the mixed liquor of water and alcohol-water3O4@SiO2Particle.Will be different
Particle is self-assembly of four kinds of magnetic pickering emulsions in oil-water interfaces.Afterwards, enzyme is fixed in lotion to measure pik woods
Emulsion intercalation method and application effect.The pickering emulsion drop stability stablized in the present invention by hydrophobically modified magnetic particle is good
It is good, it can be achieved that the cladding of a variety of enzymes, and immobilised enzymes has good heat endurance, storage stability.Four kinds of hydrophobically modified magnetic
The particle-stabilised pickering emulsion of property can realize magnetic recovery, significantly improve immobilised enzymes enzyme activity and reusing.This
The pickering emulsion cost of material of invention is low, preparation method is easily operated, there is good economic feasibility and practicality.
Beneficial effects of the present invention
The present invention utilizes hydrophobically modified Fe3O4@SiO2Particle has been prepared a variety of magnetic pickering emulsions and has been applied
In enzyme immobilization, had the following advantages compared with existing pickering emulsion：
1) present invention is to magnetic Fe3O4@SiO2Particle has carried out certain hydrophobically modified, it is stablized water-in-oil type
Pickering emulsion, and emulsion intercalation method is good (see attached drawing 1).
2) pickering emulsion for the water-in-oil type that the present invention develops can be used in enzyme immobilization system, and enzyme is coated on water
With protective enzyme in phase microenvironment, it is significantly improved (see attached drawing 2) compared with free enzyme enzyme activity.
3) the core shell structure Fe that the present invention develops3O4@SiO2Magnetic particle, easy to operate, cost is low and small toxicity, its is steady
Fixed pickering emulsion has good heat endurance and storage stability (see attached drawing 5, attached drawing 6).
4) pickering emulsion that the present invention develops is by magnetic Fe3O4@SiO2Particle forms so that pickering emulsion can be real
Existing magnetic recovery, effectively improves the reusing of immobilised enzymes (see attached drawing 7).
Brief description of the drawings
Tetra- kinds of magnetic Fes of Fig. 13O4@SiO2Particle-stabilised pickering emulsion emulsion droplet freeze after SEM and the optics of emulsion droplet show
Micro mirror figure (wherein a-e) p-Fe3O4@w-SiO2；b-f)p-Fe3O4@e-SiO2；c-g)s-Fe3O4@w-SiO2；d-h)s-Fe3O4@
e-SiO2)。
The free enzymes of Fig. 2 and four kinds of magnetic Fes3O4@SiO2The enzyme activity of particle-stabilised pickering emulsion immobilised enzymes is (wherein a)
Water/n-heptane system；B) water/n-dodecane system, oil-water ratio are respectively 10:1 and 6:1).
Fig. 3 a) oil-water ratio；b)Fe3O4@SiO2Particle load capacity is to four kinds of magnetic Fes3O4@SiO2Particle-stabilised pik woods
The influence of lotion immobilised enzymes enzyme activity.
The load capacity of the free enzymes of Fig. 4 is to magnetic p-Fe3O4@w-SiO2Particle-stabilised pickering emulsion immobilised enzymes enzyme activity
Influence.
The free enzymes of Fig. 5 and four kinds of magnetic Fes3O4@SiO2Particle-stabilised pickering emulsion immobilised enzymes is at different temperatures
The opposite enzyme activity of heating reaction 40min.
The free enzymes of Fig. 6 and four kinds of magnetic Fes3O4@SiO2Particle-stabilised pickering emulsion immobilised enzymes stores not at 4 DEG C
With the opposite enzyme activity situation of change of time.
Tetra- kinds of magnetic Fes of Fig. 73O4@SiO2Enzyme activity of the particle-stabilised pickering emulsion during recycling.
Embodiment
With reference to the drawings and specific embodiments, the present invention is described in further detail：
Hydrophobically modified magnetic Fe3O4@SiO2Particle-stabilised pickering emulsion
With magnetic Fe made from a step hydrophobically modified method3O4@SiO2Particle[3]Used as stabilizers, n-dodecane or normal heptane are made
Oil phase, Tris buffer solutions (10mM, pH 8.5) make water phase, stir above-mentioned mixed liquor using high-shear homogenizer, Fe is made3O4@SiO2
The water-in-oil type pickering emulsion that nano-particle is stablized；Wherein, magnetic Fe3O4@SiO2The volume fraction that particle accounts for water phase is 0.5-
1.5% (%, w/v), water-oil phase volume ratio are 2:1-20:1, high-shear homogenizer mixing speed is 10000-20000rpm.
Magnetic Fe3O4@SiO2Particle-stabilised pickering emulsion immobilised enzymes application
It is derived from and adds the dilution of Tris buffer solutions by enzyme solutions, makees water phase, magnetic Fe with the enzyme solutions after dilution3O4@SiO2Grain
Sub- used as stabilizers, n-dodecane make oil phase, using the above-mentioned mixed liquor of homogenizer high-speed stirred, are coated on enzyme in preparation process
In water phase, the magnetic pickering emulsion containing immobilised enzymes is obtained；Wherein, free enzyme solutions addition is 10-50 μ L, magnetic
Fe3O4@SiO2The volume fraction that particle accounts for water phase is 0.5-1.5% (%, w/v), and water-oil phase volume ratio is 2:1-20:1, it is high
Fast homogenizer speed is 10000-20000rpm.
Magnetic Fe of the present invention3O4@SiO2Particle includes p-Fe3O4@w-SiO2、p-Fe3O4@e-SiO2、s-Fe3O4@
w-SiO2、s-Fe3O4@e-SiO2Four kinds of particles.
The method of the pickering emulsion of the present invention for preparing magnetic particle stabilization, which is removed, is suitable for water/n-dodecane system
Outside, it is equally applicable to water/n-heptane system.
Free enzyme solutions of the present invention are liquid aliphatic enzyme.
Preferably, the scheme of magnetic pickering emulsion of the present invention is that water-oil phase volume ratio is 10:1, it is magnetic
Fe3O4@SiO2The volume fraction that particle accounts for water phase is 1% (%, w/v), and high-shear homogenizer mixing speed is 20000rpm.
Preferably, the free enzyme solutions addition of magnetic pickering emulsion immobilised enzymes of the present invention is 50 μ L.
Preferably, magnetic pickering emulsion of the present invention is s-Fe3O4@e-SiO2Particle-stabilised pickering emulsion.
Embodiment 1
Take 1.35g FeCl3·6H2O, 3.85g ammonium acetate and 0.4g sodium citrates are dissolved in 70mL ethylene glycol, and mixture exists
1h is stirred at 170 DEG C and forms uniform dark solution, is transferred in 100mL ptfe autoclaves, is protected at 210 DEG C afterwards
After holding 18h, room temperature is down to, is washed twice with ethanol, Magnetic Isolation, then is dried in vacuum overnight at 40 DEG C, obtains solvent heat legal system
Standby Fe3O4Particle (s-Fe3O4)。
Take Fe made from the 40mg above methods3O4It is dispersed in the mixed liquor of 16mL water and 80mL ethanol, ultrasonic 30min, adds
Enter the ammonium hydroxide of 2mL, 0.8mL ethyl orthosilicates are added dropwise afterwards, 0.16g n-octyl trimethoxy silicon is added dropwise after 5min
Alkane.Reaction system stirs 12h under 400rpm, 40 DEG C of water-baths, and Magnetic Isolation, is washed three times with ethanol, and dry 6h, obtains at 50 DEG C
s-Fe3O4@e-SiO2Particle.Take the free enzyme solutions of 20 μ L to add 980 μ L Tris buffer solutions (10mM, pH 8.5), add after mixing
Enter to being dispersed with 20mg hydrophobically modifieds s-Fe3O4@e-SiO2In the 2mL n-dodecanes of particle, high speed homogenization (20000rpm,
1min), s-Fe is obtained3O4@e-SiO2Particle-stabilised pickering emulsion immobilised enzymes system.
400mmol L will be contained-1N-caproic acid and 400mmol L-1The 2mL n-dodecane solution of n-hexyl alcohol is added to cladding
There is the catalytic effect that immobilised enzymes is measured in the pickering emulsion of enzyme.Enzymic catalytic reaction is in 37 DEG C, the shaking bath of 200rpm
40min is carried out, takes 1 μ L product gas chromatographic analysis results.The results show：s-Fe3O4@e-SiO2Particle-stabilised pik woods
The enzyme activity of lotion immobilised enzymes is 22.28U mL-1, it is significantly improved compared with free enzyme enzyme activity, about 4 times of free enzyme activity
(see attached drawing 2,3).
Embodiment 2
Take 1.35g FeCl3·6H2O, 3.85g ammonium acetate and 0.4g sodium citrates are dissolved in 70mL ethylene glycol, and mixture exists
1h is stirred at 170 DEG C and forms uniform dark solution, is transferred in 100mL ptfe autoclaves, is protected at 210 DEG C afterwards
After holding 18h, room temperature is down to, is washed twice with ethanol, Magnetic Isolation, then is dried in vacuum overnight at 40 DEG C, obtains solvent heat legal system
Standby Fe3O4Particle (s-Fe3O4)。
Take Fe made from the 40mg above methods3O4It is dispersed in the mixed liquor of 16mL water and 80mL ethanol, ultrasonic 30min, adds
Enter the ammonium hydroxide of 2mL, 0.8mL ethyl orthosilicates are added dropwise afterwards, 0.16g n-octyl trimethoxy silicon is added dropwise after 5min
Alkane.Reaction system stirs 12h under 400rpm, 40 DEG C of water-baths, and Magnetic Isolation, is washed three times with ethanol, and dry 6h, obtains at 50 DEG C
s-Fe3O4@e-SiO2Particle.Take the free enzyme solutions of 20 μ L to add 180 μ L Tris buffer solutions (10mM, pH 8.5), add after mixing
Enter to being dispersed with 20mg hydrophobically modifieds s-Fe3O4@e-SiO2In the 2mL n-dodecanes of particle, high speed homogenization (20000rpm,
1min), s-Fe is obtained3O4@e-SiO2Particle-stabilised pickering emulsion immobilised enzymes system.
400mmol L will be contained-1N-caproic acid and 400mmol L-1The 2mL n-dodecane solution of n-hexyl alcohol is added to cladding
There is the catalytic effect that immobilised enzymes is measured in the pickering emulsion of enzyme.Enzymic catalytic reaction is in 37 DEG C, the shaking bath of 200rpm
40min is carried out, takes 1 μ L product gas chromatographic analysis results.The results show：s-Fe3O4@e-SiO2Particle-stabilised pik woods
The enzyme activity of lotion immobilised enzymes is 24.17U mL-1, it is significantly improved compared with free enzyme enzyme activity, about 4 times of free enzyme activity
(see attached drawing 2,3).
Embodiment 3
Take 1.35g FeCl3·6H2O, 3.85g ammonium acetate and 0.4g sodium citrates are dissolved in 70mL ethylene glycol, and mixture exists
1h is stirred at 170 DEG C and forms uniform dark solution, is transferred in 100mL ptfe autoclaves, is protected at 210 DEG C afterwards
After holding 18h, room temperature is down to, is washed twice with ethanol, Magnetic Isolation, then is dried in vacuum overnight at 40 DEG C, obtains solvent heat legal system
Standby Fe3O4Particle (s-Fe3O4)。
Take Fe made from the 40mg above methods3O4It is dispersed in the mixed liquor of 16mL water and 80mL ethanol, ultrasonic 30min, adds
Enter the ammonium hydroxide of 2mL, 0.8mL ethyl orthosilicates are added dropwise afterwards, 0.16g n-octyl trimethoxy silicon is added dropwise after 5min
Alkane.Reaction system stirs 12h under 400rpm, 40 DEG C of water-baths, and Magnetic Isolation, is washed three times with ethanol, and dry 6h, obtains at 50 DEG C
s-Fe3O4@e-SiO2Particle.Take the free enzyme solutions of 20 μ L to add 80 μ L Tris buffer solutions (10mM, pH 8.5), added after mixing
To being dispersed with 20mg hydrophobically modifieds s-Fe3O4@e-SiO2In the 2mL n-dodecanes of particle, high speed homogenization (20000rpm, 1min),
Obtain s-Fe3O4@e-SiO2Particle-stabilised pickering emulsion immobilised enzymes system.
400mmol L will be contained-1N-caproic acid and 400mmol L-1The 2mL n-dodecane solution of n-hexyl alcohol is added to cladding
There is the catalytic effect that immobilised enzymes is measured in the pickering emulsion of enzyme.Enzymic catalytic reaction is in 37 DEG C, the shaking bath of 200rpm
40min is carried out, takes 1 μ L product gas chromatographic analysis results.The results show：s-Fe3O4@e-SiO2Particle-stabilised pik woods
The enzyme activity of lotion immobilised enzymes is 22.56U mL-1, it is significantly improved compared with free enzyme enzyme activity, about 4 times of free enzyme activity
(see attached drawing 2,3).
Embodiment 4
First prepare three kinds of solution：0.5mo/L FeSO4With 0.2mol/L HCl mixed liquors, 1mol/L FeCl3With 0.2mol/
The mixed liquor and 1.5mol/L NaOH of L HCl.100mL NaOH (1.5mol/L) are added in four-hole bottle afterwards, in 80 DEG C of water
The lower heating of bath, is added dropwise 10mL FeSO4And 10mL FeCl3Mixed liquor, reaction system is in N2Under the conditions of react 1h, when mixed
After the blackening of zoarium system, cooling continued mechanical stirring (460-470rpm), Magnetic Isolation, is washed with water three times, ethanol washes one afterwards
Time, obtain the Fe of coprecipitation preparation3O4Particle (p-Fe3O4)。
Take Fe made from the 40mg above methods3O4It is dispersed in the mixed liquor of 16mL water and 80mL ethanol, ultrasonic 30min, adds
Enter the ammonium hydroxide of 2mL, 0.8mL ethyl orthosilicates are added dropwise afterwards, 0.16g n-octyl trimethoxy silicon is added dropwise after 5min
Alkane.Reaction system stirs 12h under 400rpm, 40 DEG C of water-baths, and Magnetic Isolation, is washed three times with ethanol, and dry 6h, obtains at 50 DEG C
p-Fe3O4@e-SiO2Particle.Take the free enzyme solutions of 20 μ L to add 180 μ L Tris buffer solutions (10mM, pH 8.5), add after mixing
Enter to being dispersed with 5mg hydrophobically modifieds p-Fe3O4@e-SiO2In the 2mL n-dodecanes of particle, high speed homogenization (10000rpm,
1min), p-Fe is obtained3O4@e-SiO2Particle-stabilised pickering emulsion immobilised enzymes system.
400mmol L will be contained-1N-caproic acid and 400mmol L-1The 2mL n-dodecane solution of n-hexyl alcohol is added to cladding
There is the catalytic effect that immobilised enzymes is measured in the pickering emulsion of enzyme.Enzymic catalytic reaction is in 37 DEG C, the shaking bath of 200rpm
40min is carried out, takes 1 μ L product gas chromatographic analysis results.The results show：p-Fe3O4@e-SiO2Particle-stabilised pik woods
The enzyme activity of lotion immobilised enzymes is 22.47U mL-1, it is significantly improved compared with free enzyme enzyme activity, about 4 times of free enzyme activity
(see attached drawing 2,3).
Embodiment 5
First prepare three kinds of solution：0.5mo/L FeSO4With 0.2mol/L HCl mixed liquors, 1mol/L FeCl3With 0.2mol/
The mixed liquor and 1.5mol/L NaOH of L HCl.100mL NaOH (1.5mol/L) are added in four-hole bottle afterwards, in 80 DEG C of water
The lower heating of bath, is added dropwise 10mL FeSO4And 10mL FeCl3Mixed liquor, reaction system is in N2Under the conditions of react 1h, when mixed
After the blackening of zoarium system, cooling continued mechanical stirring (460-470rpm), Magnetic Isolation, is washed with water three times, ethanol washes one afterwards
Time, obtain the Fe of coprecipitation preparation3O4Particle (p-Fe3O4)。
Take Fe made from the 40mg above methods3O4It is dispersed in the mixed liquor of 16mL water and 80mL ethanol, ultrasonic 30min, adds
Enter the ammonium hydroxide of 2mL, 0.8mL ethyl orthosilicates are added dropwise afterwards, 0.16g n-octyl trimethoxy silicon is added dropwise after 5min
Alkane.Reaction system stirs 12h under 400rpm, 40 DEG C of water-baths, and Magnetic Isolation, is washed three times with ethanol, and dry 6h, obtains at 50 DEG C
p-Fe3O4@e-SiO2Particle.Take the free enzyme solutions of 20 μ L to add 180 μ L Tris buffer solutions (10mM, pH 8.5), add after mixing
Enter to being dispersed with 25mg hydrophobically modifieds p-Fe3O4@e-SiO2In the 2mL n-dodecanes of particle, high speed homogenization (20000rpm,
1min), p-Fe is obtained3O4@e-SiO2Particle-stabilised pickering emulsion immobilised enzymes system.
400mmol L will be contained-1N-caproic acid and 400mmol L-1The 2mL n-dodecane solution of n-hexyl alcohol is added to cladding
There is the catalytic effect that immobilised enzymes is measured in the pickering emulsion of enzyme.Enzymic catalytic reaction is in 37 DEG C, the shaking bath of 200rpm
40min is carried out, takes 1 μ L product gas chromatographic analysis results.The results show：p-Fe3O4@e-SiO2Particle-stabilised pik woods
The enzyme activity of lotion immobilization immobilised enzymes is 22.85U mL-1, it is significantly improved compared with free enzyme enzyme activity, about free enzyme
4 times (see attached drawings 2,3) living.
Embodiment 6
First prepare three kinds of solution：0.5mo/L FeSO4With 0.2mol/L HCl mixed liquors, 1mol/L FeCl3With 0.2mol/
The mixed liquor and 1.5mol/L NaOH of L HCl.100mL NaOH (1.5mol/L) are added in four-hole bottle afterwards, in 80 DEG C of water
The lower heating of bath, is added dropwise 10mL FeSO4And 10mL FeCl3Mixed liquor, reaction system is in N2Under the conditions of react 1h, when mixed
After the blackening of zoarium system, cooling continued mechanical stirring (460-470rpm), Magnetic Isolation, is washed with water three times, ethanol washes one afterwards
Time, obtain the Fe of coprecipitation preparation3O4Particle (p-Fe3O4)。
Take Fe made from the 40mg above methods3O4It is dispersed in the mixed liquor of 16mL water and 80mL ethanol, ultrasonic 30min, adds
Enter the ammonium hydroxide of 2mL, 0.8mL ethyl orthosilicates are added dropwise afterwards, 0.16g n-octyl trimethoxy silicon is added dropwise after 5min
Alkane.Reaction system stirs 12h under 400rpm, 40 DEG C of water-baths, and Magnetic Isolation, is washed three times with ethanol, and dry 6h, obtains at 50 DEG C
p-Fe3O4@e-SiO2Particle.Take the free enzyme solutions of 20 μ L to add 180 μ L Tris buffer solutions (10mM, pH 8.5), add after mixing
Enter to being dispersed with 20mg hydrophobically modifieds p-Fe3O4@e-SiO2In the 2mL n-dodecanes of particle, high speed homogenization (15000rpm,
1min), p-Fe is obtained3O4@e-SiO2Particle-stabilised pickering emulsion immobilised enzymes system.
400mmol L will be contained-1N-caproic acid and 400mmol L-1The 2mL n-dodecane solution of n-hexyl alcohol is added to cladding
There is the catalytic effect that immobilised enzymes is measured in the pickering emulsion of enzyme.Enzymic catalytic reaction is in 37 DEG C, the shaking bath of 200rpm
40min is carried out, takes 1 μ L product gas chromatographic analysis results.The results show：p-Fe3O4@e-SiO2Particle-stabilised pik woods
The enzyme activity of lotion immobilization immobilised enzymes is 23.47U mL-1, it is significantly improved compared with free enzyme enzyme activity, about free enzyme
4 times (see attached drawings 2,3) living.
Embodiment 7
Take 1.35g FeCl3·6H2O, 3.85g ammonium acetate and 0.4g sodium citrates are dissolved in 70mL ethylene glycol, and mixture exists
1h is stirred at 170 DEG C and forms uniform dark solution, is transferred in 100mL ptfe autoclaves, is protected at 210 DEG C afterwards
After holding 18h, room temperature is down to, is washed twice with ethanol, Magnetic Isolation, then is dried in vacuum overnight at 40 DEG C, obtains solvent heat legal system
Standby Fe3O4Particle (s-Fe3O4)。
Take Fe made from the 40mg above methods3O4Be dispersed in 16mL water, ultrasonic 30min, add the ammonium hydroxide of 2mL, afterwards by
0.8mL ethyl orthosilicates are added dropwise to, 0.16g n-octyl trimethoxy silanes are added dropwise after 5min.Reaction system in 400rpm,
12h is stirred under 40 DEG C of water-baths, Magnetic Isolation, is washed three times with ethanol, and dry 6h, obtains s-Fe at 50 DEG C3O4@w-SiO2Particle.Take
The free enzyme solutions of 20 μ L add 180 μ L Tris buffer solutions (10mM, pH 8.5), are added to after mixing and are dispersed with that 20mg is hydrophobic to be changed
Property s-Fe3O4@w-SiO2In the 2mL n-dodecanes of particle, high speed homogenization (20000rpm, 1min), obtains
s-Fe3O4@w-SiO2Particle-stabilised pickering emulsion immobilised enzymes system.
400mmol L will be contained-1N-caproic acid and 400mmol L-1The 2mL n-dodecane solution of n-hexyl alcohol is added to cladding
There is the catalytic effect that immobilised enzymes is measured in the pickering emulsion of enzyme.Enzymic catalytic reaction is in 37 DEG C, the shaking bath of 200rpm
40min is carried out, takes 1 μ L product gas chromatographic analysis results.The results show：s-Fe3O4@w-SiO2Particle-stabilised pik woods
The enzyme activity of lotion immobilised enzymes is 15.68U mL-1, it is significantly improved compared with free enzyme enzyme activity, about 3 times of free enzyme activity
(see attached drawing 2,3).
Embodiment 8
First prepare three kinds of solution：0.5mo/L FeSO4With 0.2mol/L HCl mixed liquors, 1mol/L FeCl3With 0.2mol/
The mixed liquor and 1.5mol/L NaOH of L HCl.100mL NaOH (1.5mol/L) are added in four-hole bottle afterwards, in 80 DEG C of water
The lower heating of bath, is added dropwise 10mL FeSO4And 10mL FeCl3Mixed liquor, reaction system is in N2Under the conditions of react 1h, when mixed
After the blackening of zoarium system, cooling continued mechanical stirring (460-470rpm), Magnetic Isolation, is washed with water three times, ethanol washes one afterwards
Time, obtain the Fe of coprecipitation preparation3O4Particle (p-Fe3O4)。
Take Fe made from the 40mg above methods3O4Be dispersed in 16mL water, ultrasonic 30min, add the ammonium hydroxide of 2mL, afterwards by
0.8mL ethyl orthosilicates are added dropwise to, 0.16g n-octyl trimethoxy silanes are added dropwise after 5min.Reaction system in 400rpm,
12h is stirred under 40 DEG C of water-baths, Magnetic Isolation, is washed three times with ethanol, and dry 6h, obtains p-Fe at 50 DEG C3O4@w-SiO2Particle.Take
The free enzyme solutions of 20 μ L add 180 μ L Tris buffer solutions (10mM, pH 8.5), are added to after mixing and are dispersed with that 20mg is hydrophobic to be changed
Property p-Fe3O4@w-SiO2In the 2mL n-dodecanes of particle, high speed homogenization (20000rpm, 1min), obtains p-Fe3O4@w-SiO2
Particle-stabilised pickering emulsion immobilised enzymes system.
400mmol L will be contained-1N-caproic acid and 400mmol L-1The 2mL n-dodecane solution of n-hexyl alcohol is added to cladding
There is the catalytic effect that immobilised enzymes is measured in the pickering emulsion of enzyme.Enzymic catalytic reaction is in 37 DEG C, the shaking bath of 200rpm
40min is carried out, takes 1 μ L product gas chromatographic analysis results.The results show：p-Fe3O4@w-SiO2Particle-stabilised pik woods
The enzyme activity of lotion immobilised enzymes is 15.32U mL-1, it is significantly improved compared with free enzyme enzyme activity, about 3 times of free enzyme activity
(see attached drawing 4).
Embodiment 9
First prepare three kinds of solution：0.5mo/L FeSO4With 0.2mol/L HCl mixed liquors, 1mol/L FeCl3With 0.2mol/
The mixed liquor and 1.5mol/L NaOH of L HCl.100mL NaOH (1.5mol/L) are added in four-hole bottle afterwards, in 80 DEG C of water
The lower heating of bath, is added dropwise 10mL FeSO4And 10mL FeCl3Mixed liquor, reaction system is in N2Under the conditions of react 1h, when mixed
After the blackening of zoarium system, cooling continued mechanical stirring (460-470rpm), Magnetic Isolation, is washed with water three times, ethanol washes one afterwards
Time, obtain the Fe of coprecipitation preparation3O4Particle (p-Fe3O4)。
Take Fe made from the 40mg above methods3O4Be dispersed in 16mL water, ultrasonic 30min, add the ammonium hydroxide of 2mL, afterwards by
0.8mL ethyl orthosilicates are added dropwise to, 0.16g n-octyl trimethoxy silanes are added dropwise after 5min.Reaction system in 400rpm,
12h is stirred under 40 DEG C of water-baths, Magnetic Isolation, is washed three times with ethanol, and dry 6h, obtains p-Fe at 50 DEG C3O4@w-SiO2Particle.Take
The free enzyme solutions of 10 μ L add 190 μ L Tris buffer solutions (10mM, pH 8.5), are added to after mixing and are dispersed with that 20mg is hydrophobic to be changed
Property p-Fe3O4@w-SiO2In the 2mL n-dodecanes of particle, high speed homogenization (20000rpm, 1min), obtains p-Fe3O4@w-SiO2
Particle-stabilised pickering emulsion immobilised enzymes system.
400mmol L will be contained-1N-caproic acid and 400mmol L-1The 2mL n-dodecane solution of n-hexyl alcohol is added to cladding
There is the catalytic effect that immobilised enzymes is measured in the pickering emulsion of enzyme.Enzymic catalytic reaction is in 37 DEG C, the shaking bath of 200rpm
40min is carried out, takes 1 μ L product gas chromatographic analysis results.The results show：p-Fe3O4@w-SiO2Particle-stabilised pik woods
The enzyme activity of lotion immobilised enzymes is 10.50U mL-1, about free enzyme activity 2 times (see attached drawings 4).
Embodiment 10
First prepare three kinds of solution：0.5mo/L FeSO4With 0.2mol/L HCl mixed liquors, 1mol/L FeCl3With 0.2mol/
The mixed liquor and 1.5mol/L NaOH of L HCl.100mL NaOH (1.5mol/L) are added in four-hole bottle afterwards, in 80 DEG C of water
The lower heating of bath, is added dropwise 10mL FeSO4And 10mL FeCl3Mixed liquor, reaction system is in N2Under the conditions of react 1h, when mixed
After the blackening of zoarium system, cooling continued mechanical stirring (460-470rpm), Magnetic Isolation, is washed with water three times, ethanol washes one afterwards
Time, obtain the Fe of coprecipitation preparation3O4Particle (p-Fe3O4)。
Take Fe made from the 40mg above methods3O4Be dispersed in 16mL water, ultrasonic 30min, add the ammonium hydroxide of 2mL, afterwards by
0.8mL ethyl orthosilicates are added dropwise to, 0.16g n-octyl trimethoxy silanes are added dropwise after 5min.Reaction system in 400rpm,
12h is stirred under 40 DEG C of water-baths, Magnetic Isolation, is washed three times with ethanol, and dry 6h, obtains p-Fe at 50 DEG C3O4@w-SiO2Particle.Take
The free enzyme solutions of 50 μ L add 150 μ L Tris buffer solutions (10mM, pH 8.5), are added to after mixing and are dispersed with that 20mg is hydrophobic to be changed
Property p-Fe3O4@w-SiO2In the 2mL n-dodecanes of particle, high speed homogenization (20000rpm, 1min), obtains p-Fe3O4@w-SiO2
Particle-stabilised pickering emulsion immobilised enzymes system.
400mmol L will be contained-1N-caproic acid and 400mmol L-1The 2mL n-dodecane solution of n-hexyl alcohol is added to cladding
There is the catalytic effect that immobilised enzymes is measured in the pickering emulsion of enzyme.Enzymic catalytic reaction is in 37 DEG C, the shaking bath of 200rpm
40min is carried out, takes 1 μ L product gas chromatographic analysis results.The results show：p-Fe3O4@w-SiO2Particle-stabilised pik woods
The enzyme activity of lotion immobilised enzymes is 19.15U mL-1, it is significantly improved compared with free enzyme enzyme activity, about 3 times of free enzyme activity
(see attached drawing 4).
Embodiment 11
First prepare three kinds of solution：0.5mo/L FeSO4With 0.2mol/L HCl mixed liquors, 1mol/L FeCl3With 0.2mol/
The mixed liquor and 1.5mol/L NaOH of L HCl.100mL NaOH (1.5mol/L) are added in four-hole bottle afterwards, in 80 DEG C of water
The lower heating of bath, is added dropwise 10mL FeSO4And 10mL FeCl3Mixed liquor, reaction system is in N2Under the conditions of react 1h, when mixed
After the blackening of zoarium system, cooling continued mechanical stirring (460-470rpm), Magnetic Isolation, is washed with water three times, ethanol washes one afterwards
Time, obtain the Fe of coprecipitation preparation3O4Particle (p-Fe3O4)。
Take Fe made from the 40mg above methods3O4Be dispersed in 16mL water, ultrasonic 30min, add the ammonium hydroxide of 2mL, afterwards by
0.8mL ethyl orthosilicates are added dropwise to, 0.16g n-octyl trimethoxy silanes are added dropwise after 5min.Reaction system in 400rpm,
12h is stirred under 40 DEG C of water-baths, Magnetic Isolation, is washed three times with ethanol, and dry 6h, obtains p-Fe at 50 DEG C3O4@w-SiO2Particle.Take
The free enzyme solutions of 20 μ L add 180 μ L Tris buffer solutions (10mM, pH 8.5), are added to after mixing and are dispersed with that 20mg is hydrophobic to be changed
Property p-Fe3O4@w-SiO2In the 2mL normal heptanes of particle, high speed homogenization (20000rpm, 1min), obtains p-Fe3O4@w-SiO2Grain
The pickering emulsion immobilised enzymes system that son is stablized.
400mmol L will be contained-1N-caproic acid and 400mmol L-1The 2mL n-heptane solutions of n-hexyl alcohol, which are added to, to be coated with
The catalytic effect of immobilised enzymes is measured in the pickering emulsion of enzyme.Enzymic catalytic reaction in 37 DEG C, the shaking bath of 200rpm into
Row 40min, takes 1 μ L product gas chromatographic analysis results.The results show：p-Fe3O4@w-SiO2Particle-stabilised pik woods breast
The enzyme activity of liquid immobilised enzymes is 8.49U mL-1(see attached drawing 2).
Embodiment 12
First prepare three kinds of solution：0.5mo/L FeSO4With 0.2mol/L HCl mixed liquors, 1mol/L FeCl3With 0.2mol/
The mixed liquor and 1.5mol/L NaOH of L HCl.100mL NaOH (1.5mol/L) are added in four-hole bottle afterwards, in 80 DEG C of water
The lower heating of bath, is added dropwise 10mL FeSO4And 10mL FeCl3Mixed liquor, reaction system is in N2Under the conditions of react 1h, when mixed
After the blackening of zoarium system, cooling continued mechanical stirring (460-470rpm), Magnetic Isolation, is washed with water three times, ethanol washes one afterwards
Time, obtain the Fe of coprecipitation preparation3O4Particle (p-Fe3O4)。
Take Fe made from the 40mg above methods3O4It is dispersed in the mixed liquor of 16mL water and 80mL ethanol, ultrasonic 30min, adds
Enter the ammonium hydroxide of 2mL, 0.8mL ethyl orthosilicates are added dropwise afterwards, 0.16g n-octyl trimethoxy silicon is added dropwise after 5min
Alkane.Reaction system stirs 12h under 400rpm, 40 DEG C of water-baths, and Magnetic Isolation, is washed three times with ethanol, and dry 6h, obtains at 50 DEG C
p-Fe3O4@e-SiO2Particle.Take the free enzyme solutions of 20 μ L to add 180 μ L Tris buffer solutions (10mM, pH 8.5), add after mixing
Enter to being dispersed with 20mg hydrophobically modifieds p-Fe3O4@e-SiO2In the 2mL normal heptanes of particle, high speed homogenization (20000rpm, 1min),
Obtain p-Fe3O4@e-SiO2Particle-stabilised pickering emulsion immobilised enzymes system.
400mmol L will be contained-1N-caproic acid and 400mmol L-1The 2mL n-heptane solutions of n-hexyl alcohol, which are added to, to be coated with
The catalytic effect of immobilised enzymes is measured in the pickering emulsion of enzyme.Enzymic catalytic reaction in 37 DEG C, the shaking bath of 200rpm into
Row 40min, takes 1 μ L product gas chromatographic analysis results.The results show：p-Fe3O4@e-SiO2Particle-stabilised pik woods breast
The enzyme activity of liquid immobilised enzymes is 23.29U mL-1, it is significantly improved compared with free enzyme enzyme activity, about 4 times of free enzyme activity
(see attached drawing 2).
Embodiment 13
Take 1.35g FeCl3·6H2O, 3.85g ammonium acetate and 0.4g sodium citrates are dissolved in 70mL ethylene glycol, and mixture exists
1h is stirred at 170 DEG C and forms uniform dark solution, is transferred in 100mL ptfe autoclaves, is protected at 210 DEG C afterwards
After holding 18h, room temperature is down to, is washed twice with ethanol, Magnetic Isolation, then is dried in vacuum overnight at 40 DEG C, obtains solvent heat legal system
Standby Fe3O4Particle (s-Fe3O4)。
Take Fe made from the 40mg above methods3O4Be dispersed in 16mL water, ultrasonic 30min, add the ammonium hydroxide of 2mL, afterwards by
0.8mL ethyl orthosilicates are added dropwise to, 0.16g n-octyl trimethoxy silanes are added dropwise after 5min.Reaction system in 400rpm,
12h is stirred under 40 DEG C of water-baths, Magnetic Isolation, is washed three times with ethanol, and dry 6h, obtains s-Fe at 50 DEG C3O4@w-SiO2Particle.Take
The free enzyme solutions of 20 μ L add 180 μ L Tris buffer solutions (10mM, pH 8.5), are added to after mixing and are dispersed with that 20mg is hydrophobic to be changed
Property s-Fe3O4@w-SiO2In the 2mL normal heptanes of particle, high speed homogenization (20000rpm, 1min), obtains s-Fe3O4@w-SiO2Grain
The pickering emulsion immobilised enzymes system that son is stablized.
400mmol L will be contained-1N-caproic acid and 400mmol L-1The 2mL n-heptane solutions of n-hexyl alcohol, which are added to, to be coated with
The catalytic effect of immobilised enzymes is measured in the pickering emulsion of enzyme.Enzymic catalytic reaction in 37 DEG C, the shaking bath of 200rpm into
Row 40min, takes 1 μ L product gas chromatographic analysis results.The results show：s-Fe3O4@w-SiO2Particle-stabilised pik woods breast
The enzyme activity of liquid immobilised enzymes is 11.13U mL-1, about free enzyme activity 2 times (see attached drawings 2).
Embodiment 14
Take 1.35g FeCl3·6H2O, 3.85g ammonium acetate and 0.4g sodium citrates are dissolved in 70mL ethylene glycol, and mixture exists
1h is stirred at 170 DEG C and forms uniform dark solution, is transferred in 100mL ptfe autoclaves, is protected at 210 DEG C afterwards
After holding 18h, room temperature is down to, is washed twice with ethanol, Magnetic Isolation, then is dried in vacuum overnight at 40 DEG C, obtains solvent heat legal system
Standby Fe3O4Particle (s-Fe3O4)。
Take Fe made from the 40mg above methods3O4It is dispersed in the mixed liquor of 16mL water and 80mL ethanol, ultrasonic 30min, adds
Enter the ammonium hydroxide of 2mL, 0.8mL ethyl orthosilicates are added dropwise afterwards, 0.16g n-octyl trimethoxy silicon is added dropwise after 5min
Alkane.Reaction system stirs 12h under 400rpm, 40 DEG C of water-baths, and Magnetic Isolation, is washed three times with ethanol, and dry 6h, obtains at 50 DEG C
s-Fe3O4@e-SiO2Particle.Take the free enzyme solutions of 20 μ L to add 180 μ L Tris buffer solutions (10mM, pH 8.5), add after mixing
Enter to being dispersed with 20mg hydrophobically modifieds s-Fe3O4@e-SiO2In the 2mL normal heptanes of particle, high speed homogenization (20000rpm, 1min),
Obtain s-Fe3O4@e-SiO2Particle-stabilised pickering emulsion immobilised enzymes system.
400mmol L will be contained-1N-caproic acid and 400mmol L-1The 2mL n-heptane solutions of n-hexyl alcohol, which are added to, to be coated with
The catalytic effect of immobilised enzymes is measured in the pickering emulsion of enzyme.Enzymic catalytic reaction in 37 DEG C, the shaking bath of 200rpm into
Row 40min, takes 1 μ L product gas chromatographic analysis results.The results show：s-Fe3O4@e-SiO2Particle-stabilised pik woods breast
The enzyme activity of liquid immobilised enzymes is 23.59U mL-1, it is significantly improved compared with free enzyme enzyme activity, about 4 times of free enzyme activity
(see attached drawing 2).
Claims (9)
1. a kind of preparation method for the water-in-oil type pickering emulsion stablized based on hydrophobically modified magnetic particle, it is characterized in that：With
Magnetic Fe made from one step hydrophobically modified method3O4@SiO2Particle used as stabilizers, n-dodecane or normal heptane make oil phase, Tris bufferings
Liquid makees water phase, stirs above-mentioned mixed liquor using high-shear homogenizer, Fe is made3O4@SiO2The water-in-oil type pik that nano-particle is stablized
Woods lotion.
2. the method as described in claim 1, it is characterized in that magnetic Fe3O4@SiO2The volume fraction that particle accounts for water phase is 0.5-
1.5% (%, w/v), water-oil phase volume ratio are 2:1-20:1.
3. the method as described in claim 1, it is characterized in that high-shear homogenizer mixing speed is 10000-20000rpm.
4. the method as described in claim 1, it is characterized in that Tris buffer solutions are 10mM, pH 8.5.
5. the according to the method described in claim 1, it is characterized in that magnetic Fe3O4@SiO2Particle includes：p-Fe3O4@w-
SiO2、p-Fe3O4@e-SiO2、s-Fe3O4@w-SiO2Or s-Fe3O4@e-SiO2。
6. according to the method described in claim 1, it is characterized in that the magnetic pickering emulsion is s-Fe3O4@e-SiO2Grain
The pickering emulsion that son is stablized.
7. the according to the method described in claim 1, it is characterized in that water-oil phase volume ratio of the magnetic pickering emulsion
For 10:1, magnetic Fe3O4@SiO2The volume fraction that particle accounts for water phase is 1% (%, w/v), and high-shear homogenizer mixing speed is
20000rpm。
8. hydrophobically modified magnetic Fe prepared by claim 13O4@SiO2Particle-stabilised pickering emulsion in immobilised enzymes application, its
It is characterized in：It is derived from and adds the dilution of Tris buffer solutions by enzyme solutions, makees water phase, magnetic Fe with the enzyme solutions after dilution3O4@SiO2Grain
Sub- used as stabilizers, n-dodecane make oil phase, using the above-mentioned mixed liquor of homogenizer high-speed stirred, are coated on enzyme in preparation process
In water phase, the magnetic pickering emulsion containing immobilised enzymes is obtained；Wherein, free enzyme solutions addition is 10-50 μ L, magnetic
Fe3O4@SiO2The volume fraction that particle accounts for water phase is 0.5-1.5% (%, w/v), and water-oil phase volume ratio is 2:1-20:1, it is high
Fast homogenizer speed is 10000-20000rpm.
9. according to the method described in claim 8, it is characterized in that the free enzyme solutions are liquid aliphatic enzyme.
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| CN108686575A (en) * | 2018-05-15 | 2018-10-23 | 江南大学 | A kind of compound emulsifying agent with magnetic responsiveness energy |
| CN109706141A (en) * | 2019-02-01 | 2019-05-03 | 中国农业科学院油料作物研究所 | A kind of immobilised enzymes pickering emulsion reaction system and its application |
| CN110373171A (en) * | 2019-07-10 | 2019-10-25 | 浙江海洋大学 | A kind of displacement of reservoir oil emulsion stable by magnetic nanoparticle |
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| CN112812760A (en) * | 2021-01-13 | 2021-05-18 | 西南石油大学 | Magnetic response Janus nanoparticle based on oil-in-water high internal phase emulsification and preparation method thereof |
| CN113105572A (en) * | 2021-03-15 | 2021-07-13 | 江苏科技大学 | Aflatoxin magnetic molecularly imprinted polymer and preparation method and application thereof |
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| CN108686575A (en) * | 2018-05-15 | 2018-10-23 | 江南大学 | A kind of compound emulsifying agent with magnetic responsiveness energy |
| CN109706141A (en) * | 2019-02-01 | 2019-05-03 | 中国农业科学院油料作物研究所 | A kind of immobilised enzymes pickering emulsion reaction system and its application |
| CN109706141B (en) * | 2019-02-01 | 2022-09-09 | 中国农业科学院油料作物研究所 | Immobilized enzyme pickering emulsion reaction system and application thereof |
| CN110373171A (en) * | 2019-07-10 | 2019-10-25 | 浙江海洋大学 | A kind of displacement of reservoir oil emulsion stable by magnetic nanoparticle |
| CN110373171B (en) * | 2019-07-10 | 2021-07-13 | 浙江海洋大学 | Oil repellent emulsion stabilized by magnetic nanoparticles |
| CN111808839A (en) * | 2020-06-10 | 2020-10-23 | 广东省测试分析研究所（中国广州分析测试中心） | Lipase interface immobilization method based on hydrophobic modified gelatin microspheres |
| CN111808839B (en) * | 2020-06-10 | 2021-05-11 | 广东省测试分析研究所（中国广州分析测试中心） | Lipase interface immobilization method based on hydrophobic modified gelatin microspheres |
| CN112812760A (en) * | 2021-01-13 | 2021-05-18 | 西南石油大学 | Magnetic response Janus nanoparticle based on oil-in-water high internal phase emulsification and preparation method thereof |
| CN112812760B (en) * | 2021-01-13 | 2022-06-10 | 西南石油大学 | Magnetic response Janus nanoparticle based on oil-in-water high internal phase emulsification and preparation method thereof |
| CN113105572A (en) * | 2021-03-15 | 2021-07-13 | 江苏科技大学 | Aflatoxin magnetic molecularly imprinted polymer and preparation method and application thereof |
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Application publication date: 20180420 |