CN104525092A - Preparation method and product of superparamagnetism nano particles - Google Patents

Preparation method and product of superparamagnetism nano particles Download PDF

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CN104525092A
CN104525092A CN201410787362.XA CN201410787362A CN104525092A CN 104525092 A CN104525092 A CN 104525092A CN 201410787362 A CN201410787362 A CN 201410787362A CN 104525092 A CN104525092 A CN 104525092A
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preparation
nano particle
magnetic nanoparticle
magnetic
alkali lye
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CN104525092B (en
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杨海
杨勇
黄燕婷
周坤
向雯琦
金伟
杨祥良
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Guangna Dakang (Guangzhou) Biotechnology Co., Ltd
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WUHAN BEST HEALTH BIOTECHNOLOGY CO Ltd
Huazhong University of Science and Technology
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/02—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material
    • B01J20/06—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material comprising oxides or hydroxides of metals not provided for in group B01J20/04
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28002—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their physical properties
    • B01J20/28009—Magnetic properties
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/3085—Chemical treatments not covered by groups B01J20/3007 - B01J20/3078
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C07K1/16—Extraction; Separation; Purification by chromatography
    • C07K1/22—Affinity chromatography or related techniques based upon selective absorption processes
    • H—ELECTRICITY
    • H01—ELECTRIC ELEMENTS
    • H01F—MAGNETS; INDUCTANCES; TRANSFORMERS; SELECTION OF MATERIALS FOR THEIR MAGNETIC PROPERTIES
    • H01F1/00—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties
    • H01F1/01—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties of inorganic materials
    • H01F1/03—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties of inorganic materials characterised by their coercivity
    • H01F1/032—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties of inorganic materials characterised by their coercivity of hard-magnetic materials
    • H01F1/04—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties of inorganic materials characterised by their coercivity of hard-magnetic materials metals or alloys
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2220/00—Aspects relating to sorbent materials
    • B01J2220/40—Aspects relating to the composition of sorbent or filter aid materials
    • B01J2220/48—Sorbents characterised by the starting material used for their preparation
    • B01J2220/4806—Sorbents characterised by the starting material used for their preparation the starting material being of inorganic character

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  • General Health & Medical Sciences (AREA)
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  • Proteomics, Peptides & Aminoacids (AREA)
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  • Compounds Of Iron (AREA)

Abstract

The invention discloses a preparation method of water-soluble superparamagnetism nano particles with high adsorption capacity for proteins. The method is characterized in that aluminum ions of an appropriate amount are added in the process for preparing ferriferrous oxide in a co-precipitation method, Fe<3+>, Al<3+> and Fe<2+> are co-precipitated under the effect of alkaline solution, and the superparamagnetism ferric oxide nano particles with a great amount of positive charges are generated. Compared with the ferriferous oxide nano particles prepared in a traditional co-precipitation method, the surface of the ferric oxide nano particles prepared in the method is provided with more positive charges, so that the stability in an aqueous solution is better, and the adsorption capacity for the natural proteins with negative charges is higher.

Description

Preparation method of a kind of superparamagnetic nano particle and products thereof
Technical field
The invention belongs to field of nanometer material technology, more specifically, relate to preparation method of a kind of superparamagnetic nano particle and products thereof.
Background technology
Magnetic nanoparticle is that one is in nano level magnetic material, possesses quantum size effect, skin effect, small-size effect and macro quanta tunnel effect etc.Magnetic nanoparticle has good magnetic conductance tropism, biocompatibility and biodegradability etc., can in conjunction with multiple biologically functional molecule, as enzyme, DNA, protein etc., in DNA separation and purification, magnetic targeted administration, medical science detection, diagnosis, gene therapy, cell separation, immunoassay, immobilised enzymes and protein etc., have a wide range of applications.
The preparation of magnetic nanoparticle, generally has radiation polymerization, thermal curing method and coprecipitation, and wherein coprecipitation has the advantages such as preparation technology is simple, cost is low, synthesis cycle is short.But SPIO nanoparticle surface band positive charge prepared by this coprecipitation is lower, and the native protein overwhelming majority under physiological condition is with negative electrical charge, the adsorption capacity of super-paramagnetism nano grain to protein that this makes coprecipitation prepare is more weak, is unfavorable for the preparation of large biological molecule coated magnetic particulate.
Summary of the invention
For above defect or the Improvement requirement of prior art, the invention provides preparation method of a kind of superparamagnetic nano particle and products thereof, its object is to prepare in the process of tri-iron tetroxide in coprecipitation increase appropriate aluminium ion, thus the SPIO nanoparticle generated with a large amount of positive charge, solving coprecipitation thus, to prepare SPIO nanoparticle surface band positive charge low, the technical problem not high to the native protein adsorption capacity with negative electrical charge.
For achieving the above object, according to one aspect of the present invention, provide a kind of preparation method of superparamagnetic nano particle, comprise the following steps:
(1) under starvation condition, by Fe 3+, Al 3+, Fe 2+be dispersed in water and form precursor solution, make Fe 3+concentration between 0.05mol/L to 0.5mol/L, Fe 3+and Fe 2+mol ratio be 2:1, Fe 3+and Al 3+mol ratio between 15:1 to 4:1;
(2) under starvation condition, by the precursor solution Keep agitation obtained in step (1), holding temperature 30 DEG C to 60 DEG C, and slowly dropping alkali lye regulates solution ph to be more than or equal to 9, forms suspension;
(3) under starvation condition, by the suspension Keep agitation obtained in step (2), and be warming up to 60 DEG C to 90 DEG C, reaction 0.5 is little of 2 hours, and cooling is precipitation and separation also, obtains described magnetic nanoparticle after cleaning.。
Preferably, described preparation method, in its step (2), mixing speed is 800rpm to 2000rpm, and the rate of addition of alkali lye is 0.5mL/min to 4mL/min.
Preferably, described preparation method, its step (2) described alkali lye is the sodium carbonate liquor of pH >=9.5, ammonia spirit or sodium hydroxide solution.
Preferably, described preparation method, its step (2) and step (3) described stirring are mechanical agitation and/or ultrasonic vibration.
According to another aspect of the present invention, provide the magnetic nanoparticle that a kind of described preparation method prepares, described magnetic nanoparticle has superparamagnetism.
Preferably, described magnetic nanoparticle, the Zeta potential of magnetic nanoparticle described in it is between+25mV to+41mV.
In general, the above technical scheme conceived by the present invention compared with prior art, can obtain following beneficial effect:
(1) preparation method of nano particle provided by the invention, on the basis of the preparation method of conventional coprecipitation, by adding appropriate Al 3+, form special colloid reaction system, thus preparation has the nano particle of superparamagnetism, technique is simple, cost is low, synthesis cycle is short.
(2) according to nano particle prepared by nanometer grain preparation method provided by the invention, have superparamagnetism, surface is with a large amount of positive charge simultaneously, and its Zeta potential is between+25mV to+41mV, to the native protein with negative electrical charge, high adsorption capacity.The water-soluble SPIO material of this height absorption is conducive to forming more stable albumen hat (protein corona) parcel on nanoparticle surface, thus improve stability and the biocompatibility of the iron oxide nano-granule aqueous solution, the biomedical sectors such as medical image, Magneto separate reagent, drug carrier material, immune diagnostic reagent can be widely used in.
Accompanying drawing explanation
Fig. 1 is transmission electron microscope (TEM) photo of magnetic nano particle prepared by the inventive method;
Fig. 2 is the hydration grain size distribution (DLS method) of magnetic nano particle prepared by the inventive method;
Fig. 3 is the magnetic hysteresis loop (VSM method) of magnetic nano particle prepared by the inventive method;
Fig. 4 is that the surface charge (Zeta potential) of magnetic nano particle MNP2 prepared by the magnetic nano particle MNP1 for preparing of embodiment 5 traditional co-precipitation method and the inventive method contrasts;
Fig. 5 is that the magnetic nano particle MNP2 for preparing of the magnetic nano particle MNP1 for preparing of embodiment 5 traditional co-precipitation method and the inventive method is to the contrast (thermogravimetric curve) of bovine serum albumin(BSA) BSA adsorbance;
Fig. 6 be the magnetic nano particle MNP2 for preparing of magnetic nano particle MNP1, the inventive method prepared by embodiment 7 traditional co-precipitation method and after add Al 3+surface charge (Zeta potential) contrast of the magnetic nano particle MNP3 of preparation;
Fig. 7 be the magnetic nano particle MNP2 for preparing of magnetic nano particle MNP1, the inventive method that embodiment 7pH 5.0 cushioning liquid is prepared traditional co-precipitation method and after add Al 3+sedimentation situation after the magnetic nano particle MNP3 effect of preparation;
Fig. 8 be the magnetic nano particle MNP2 for preparing of magnetic nano particle MNP1, the inventive method prepared by embodiment 7 traditional co-precipitation method and after add Al 3+the magnetic nano particle MNP3 of preparation is to the contrast (UV absorptiometry) of bovine serum albumin(BSA) BSA adsorbance.
Detailed description of the invention
In order to make object of the present invention, technical scheme and advantage clearly understand, below in conjunction with drawings and Examples, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.In addition, if below in described each embodiment of the present invention involved technical characteristic do not form conflict each other and just can mutually combine.
The preparation method of superparamagnetic nano particle provided by the invention, comprises the following steps:
(1) under starvation condition, by Fe 3+, Al 3+, Fe 2+be dispersed in water and form precursor solution, make Fe 3+concentration between 0.05mol/L to 0.5mol/L, Fe 3+and Fe 2+mol ratio be 2:1, Fe 3+and Al 3+mol ratio between 15:1 to 4:1.
(2) under starvation condition, by the precursor solution Keep agitation obtained in step (1), holding temperature 30 DEG C to 60 DEG C, and slowly dropping alkali lye regulates solution ph to be more than or equal to 9, forms suspension; Preferably, mixing speed is 800rpm to 2000rpm, and the rate of addition of alkali lye is 0.5mL/min to 4mL/min.Described alkali lye is the sodium carbonate liquor of pH >=9.5, ammonia spirit or sodium hydroxide solution.For make product disperse evenly, also can add citrate ion in alkali lye.
(3) under starvation condition, by the suspension Keep agitation obtained in step (2), and be warming up to 60 DEG C to 90 DEG C, reaction 0.5 is little of 2 hours, and cooling is precipitation and separation also, obtains described magnetic nanoparticle after cleaning.
Step (2) and step (3) described stirring are mechanical agitation and/or ultrasonic vibration.
According to method provided by the invention, the magnetic nanoparticle of preparation, has superparamagnetism, and its Zeta potential is between+25mV to+41mV.Compare magnetic nanoparticle prepared by existing chemical coprecipitation technique, magnetic nanoparticle surface prepared by the present invention, with more positive charge, is more conducive to adsorb electronegative albumen.Magnetic nano particle prepared by the inventive method, as shown in Figure 1, as shown in Figure 2, its magnetic hysteresis loop (VSM method) as shown in Figure 3 for its hydration grain size distribution (DLS method) for its transmission electron microscope (TEM) photo.
Be below embodiment:
Embodiment 1
A preparation method for superparamagnetic nano particle, comprises the following steps:
(1) get 250mL ultra-pure water, logical nitrogen, after 10 minutes, adds 7.78g FeCl successively 36H 2o, 1.60g Al (NO 3) 39H 2o and 2.86g FeCl 24H 2o, by Fe 3+, Al 3+, Fe 2+be dispersed in water and form precursor solution, make Fe 3+concentration be 0.115mol/L, Fe 3+and Fe 2+mol ratio be 2:1, Fe 3+and Al 3+mol ratio be 6.4:1.
(2) under nitrogen protection, by the precursor solution Keep agitation obtained in step (1), holding temperature 40 DEG C, and drip alkali lye 100mL, alkali lye is the ammonia spirit of 2mol/L, final solution pH value >=9, forms suspension; Mixing speed 1500rpm, the rate of addition of alkali lye is 2mL/min.
(3) under nitrogen protection, by the suspension Keep agitation obtained in step (2), and be warming up to 80 DEG C, react 0.5 hour, product is cooled to room temperature, and supernatant is removed in magnetic sedimentation separation; Use 100mL ultra-pure water resuspended after cleaning 5 times with ultra-pure water, preserve in 4 DEG C of sealings.
According to the magnetic nanoparticle that described preparation method prepares, be black, energy sedimentation completely under additional magnetic fields, record average hydration particle diameter through dynamic light scattering method (DLS) and be about 180nm, Zeta potential is+36 ± 5mV.Characterize through transmission electron microscope TEM, individual particle particle diameter is 15 to 20nm.
Embodiment 2
(1) get 250mL ultra-pure water, logical nitrogen, after 10 minutes, adds 20.27g FeCl successively 36H 2o, 7.03g Al (NO 3) 39H 2o and 7.46g FeCl 24H 2o, by Fe 3+, Al 3+, Fe 2+be dispersed in water and form precursor solution, make Fe 3+concentration be 0.3mol/L, Fe 3+and Fe 2+mol ratio be 2:1, Fe 3+and Al 3+mol ratio be 4:1.
(2) under nitrogen protection, by the precursor solution Keep agitation obtained in step (1), holding temperature 30 DEG C, and drip alkali lye 200mL, alkali lye is the NaOH solution of 1mol/L, final solution pH value >=9, forms suspension; Mixing speed 2000rpm, the rate of addition of alkali lye is 4mL/min.While dripping alkali lye, with the supersonic generator of 400kHz, 50W, ultrasonic vibration is aided with to whole reaction vessel.
(3) under nitrogen protection, by the suspension Keep agitation obtained in step (2), and be warming up to 60 DEG C, react 1 hour, product is cooled to room temperature, and supernatant is removed in magnetic sedimentation separation; Use 100mL ultra-pure water resuspended after cleaning 5 times with ultra-pure water, preserve in 4 DEG C of sealings.
According to the magnetic nanoparticle that described preparation method prepares, be black, energy sedimentation completely under additional magnetic fields, record average hydration particle diameter through dynamic light scattering method (DLS) and be about 200nm, Zeta potential is+32 ± 4mV.Characterize through transmission electron microscope TEM, individual particle particle diameter is 15 to 20nm.
Embodiment 3
(1) get 250mL ultra-pure water, logical nitrogen, after 10 minutes, adds 33.79g FeCl successively 36H 2o, 3.13g Al (NO 3) 39H 2o and 12.43g FeCl 24H 2o, by Fe 3+, Al 3+, Fe 2+be dispersed in water and form precursor solution, make Fe 3+concentration be 0.5mol/L, Fe 3+and Fe 2+mol ratio be 2:1, Fe 3+and Al 3+mol ratio be 15:1.
(2) under nitrogen protection, by the precursor solution Keep agitation obtained in step (1), holding temperature 60 DEG C, and drip alkali lye 50mL, alkali lye is the sodium carbonate liquor of 2mol/L, final solution pH value >=9, forms suspension; Mixing speed 800rpm, the rate of addition of alkali lye is 0.5mL/min.
(3) under nitrogen protection, by the suspension Keep agitation obtained in step (2), and be warming up to 90 DEG C, react 2 hours, product is cooled to room temperature, and supernatant is removed in magnetic sedimentation separation; Use 100mL ultra-pure water resuspended after cleaning 5 times with ultra-pure water, preserve in 4 DEG C of sealings.
According to the magnetic nanoparticle that described preparation method prepares, be black, energy sedimentation completely under additional magnetic fields, record average hydration particle diameter through dynamic light scattering method (DLS) and be about 210nm, Zeta potential is+30 ± 5mV.Characterize through transmission electron microscope TEM, individual particle particle diameter is about 15 to 20nm.
Embodiment 4
(1) get 250mL ultra-pure water, logical nitrogen, after 10 minutes, adds 3.38g FeCl successively 36H 2o, 0.73g Al (NO 3) 39H 2o and 1.24g FeCl 24H 2o, by Fe 3+, Al 3+, Fe 2+be dispersed in water and form precursor solution, make Fe 3+concentration be 0.05mol/L, Fe 3+and Fe 2+mol ratio be 2:1, Fe 3+and Al 3+mol ratio be 6.4:1.
(2) under nitrogen protection, by the precursor solution Keep agitation obtained in step (1), holding temperature 40 DEG C, and drip alkali lye 100mL, alkali lye is the ammonia spirit of 2mol/L, final solution pH value >=9, forms suspension; Mixing speed 1500rpm, the rate of addition of alkali lye is 2mL/min.
(3) under nitrogen protection, by the suspension Keep agitation obtained in step (2), and be warming up to 80 DEG C, react 0.5 hour, product is cooled to room temperature, and supernatant is removed in magnetic sedimentation separation; Use 100mL ultra-pure water resuspended after cleaning 5 times with ultra-pure water, preserve in 4 DEG C of sealings.
According to the magnetic nanoparticle that described preparation method prepares, be black, energy sedimentation completely under additional magnetic fields, record average hydration particle diameter through dynamic light scattering method (DLS) and be about 180nm, Zeta potential is+30mV ± 5mV.Characterize through transmission electron microscope TEM, individual particle particle diameter is about 15 to 20nm.
The contrast of magnetic nano particle prepared by embodiment 5 and traditional co-precipitation method
Prepare magnetic nano particle MNP1 prepared by traditional co-precipitation method according to the following steps
(1) get 250mL ultra-pure water, logical nitrogen, after 10 minutes, adds 7.78g FeCl successively 36H 2o and 2.86g FeCl 24H 2o, by Fe 3+, Fe 2+be dispersed in water and form precursor solution, make Fe 3+concentration be 0.115mol/L, Fe 3+and Fe 2+mol ratio be 2:1.
(2) under nitrogen protection, by the precursor solution Keep agitation obtained in step (1), holding temperature 40 DEG C, and drip alkali lye 100mL, alkali lye is the ammonia spirit of 2mol/L, final solution pH value >=9, forms suspension; Mixing speed 1500rpm, the rate of addition of alkali lye is 2mL/min.
(3) under nitrogen protection, by the suspension Keep agitation obtained in step (2), and be warming up to 80 DEG C, react 0.5 hour, product is cooled to room temperature, and supernatant is removed in magnetic sedimentation separation; Use 100mL ultra-pure water resuspended after cleaning 5 times with ultra-pure water, preserve in 4 DEG C of sealings, be labeled as MNP1.
Prepare magnetic nano particle MNP2 prepared by the inventive method according to the following steps
(1) get 250mL ultra-pure water, logical nitrogen, after 10 minutes, adds 7.78g FeCl successively 36H 2o, 1.60g Al (NO 3) 39H 2o and 2.86g FeCl 24H 2o, by Fe 3+, Al 3+, Fe 2+be dispersed in water and form precursor solution, make Fe 3+concentration be 0.115mol/L, Fe 3+and Fe 2+mol ratio be 2:1, Fe 3+and Al 3+mol ratio be 6.4:1.
(2) under nitrogen protection, by the precursor solution Keep agitation obtained in step (1), holding temperature 40 DEG C, and drip alkali lye 100mL, alkali lye is the ammonia spirit of 2mol/L, final solution pH value >=10, forms suspension; Mixing speed 1500rpm, the rate of addition of alkali lye is 2mL/min.
(3) under nitrogen protection, by the suspension Keep agitation obtained in step (2), and be warming up to 80 DEG C, react 0.5 hour, product is cooled to room temperature, and supernatant is removed in magnetic sedimentation separation; Use 100mL ultra-pure water resuspended after cleaning 5 times with ultra-pure water, preserve in 4 DEG C of sealings, be labeled as MNP2.
Both Zeta potential results as shown in Figure 4, magnetic nano particle Zeta potential prepared by traditional co-precipitation method is+24 ± 5mV, and magnetic nano particle Zeta potential prepared by the inventive method is+36 ± 5mV, its surface institute is positively charged more, is more conducive to adsorbing electronegative albumen.
Protein adsorption quantitative determination is carried out as follows.
(1) get the centrifuge tube of 4 2mL, two magnetic nano particle MNP1 adding 3mg traditional co-precipitation method and prepare, another two magnetic nano particle MNP2 adding 3mg the inventive method and prepare, clean 3 times with 1mL ultra-pure water respectively, outwell supernatant.
(2) centrifuge tube of 2 2mL is got, take 30mg BSA respectively, be dissolved in 1mL ultra-pure water, one is mixed into MNP1+BSA group with MNP1, another and MNP2 are mixed into MNP2+BSA group, it is MNP1 group and MNP2 group that remaining MNP1 and MNP2 adds 1mL ultra-pure water respectively, and four pipes are mixed 2h on blood mixer.
(3) all samples is carried out magnetic sedimentation, remove supernatant, then clean 1 time with 1mL ultra-pure water, remove supernatant, sample is placed on-20 DEG C of pre-freezes and spends the night, then freeze-drying in freeze dryer.
(4) analyze sample with thermogravimetric analyzer (Pyris1TGA, platinum-Ai Ermo instrument (Shanghai) Co., Ltd.), condition is 50-800 DEG C, heating rate 10 DEG C/min.
As shown in Figure 5, magnetic nano particle MNP1 prepared by traditional co-precipitation method and magnetic nano particle MNP2 weightlessness about 5% prepared by the inventive method, so physical absorption water accounts for 5% to thermogravimetric result.MNP1+BSA weightlessness about 16%, MNP2+BSA weightlessness about 24%, deduct physical absorption water proportion, so the BSA adsorbance of MNP1 and MNP2 is respectively 11% and 19%, the magnetic nano particle height about 72.7% that the BSA adsorbance of magnetic nano particle prepared by the inventive method is prepared than traditional co-precipitation method.
The impact of embodiment 6 aluminium ion addition
(1) get 250mL ultra-pure water, logical nitrogen, after 10 minutes, adds 7.78g FeCl successively 36H 2o, 3.20g Al (NO 3) 39H 2o and 2.86g FeCl 24H 2o, by Fe 3+, Al 3+, Fe 2+be dispersed in water and form precursor solution, make Fe 3+concentration be 0.115mol/L, Fe 3+and Fe 2+mol ratio be 2:1, Fe 3+and Al 3+mol ratio be 3.2:1.
(2) under nitrogen protection, by the precursor solution Keep agitation obtained in step (1), holding temperature 40 DEG C, and drip alkali lye 100mL, alkali lye is the ammonia spirit of 2mol/L, final solution pH value >=9, forms suspension; Mixing speed 1500rpm, the rate of addition of alkali lye is 2mL/min.
(3) under nitrogen protection, by the suspension Keep agitation obtained in step (2), and be warming up to 80 DEG C, react 0.5 hour, product is cooled to room temperature, and supernatant is removed in magnetic sedimentation separation; Use 100mL ultra-pure water resuspended after cleaning 5 times with ultra-pure water, preserve in 4 DEG C of sealings.
By Al (NO 3) 39H 2the amount of O changes 3.20g into, Fe 3+and Al 3+mol ratio be about 3.2:1, all the other are tested according to the step that embodiment 1 is identical, and result products therefrom is brown, and can not magnetic sedimentation completely, show that the aluminium ion of excessive concentrations can hinder the generation of SPIO nanoparticle.Only have Fe by the present invention 3+and Al 3+mol ratio control between 15:1 to 4:1, just can prepare high absorption super-paramagnetism nano grain.
The impact of embodiment 7 aluminium ion addition sequence
Magnetic nano particle MNP1 prepared by traditional co-precipitation method and magnetic nano particle MNP2 prepared by the inventive method is prepared respectively by step in embodiment 5.
Al is added according to the following steps after preparation 3+the magnetic nano particle MNP3 of preparation
(1) magnetic nano particle MNP1 prepared by traditional co-precipitation method is prepared by step in embodiment 5.
(2) get MNP1 prepared by step (1), add 150mL ultra-pure water, add 1.60gAl (NO 3) 39H 2o, under nitrogen protection, Keep agitation, holding temperature 40 DEG C, and drip alkali lye 100mL, alkali lye is the ammonia spirit of 2mol/L, final solution pH value>=9, forms suspension; Mixing speed 1500rpm, the rate of addition of alkali lye is 2mL/min.
(3) under nitrogen protection, by the suspension Keep agitation obtained in step (4), and be warming up to 80 DEG C, react 0.5 hour, product is cooled to room temperature, and supernatant is removed in magnetic sedimentation separation; Use 100mL ultra-pure water resuspended after cleaning 5 times with ultra-pure water, preserve in 4 DEG C of sealings, be labeled as MNP3.
By Al (NO 3) 39H 2after the joining day of O changes into and prepares magnetic nano particle MNP1 prepared by traditional co-precipitation method, gained SPIO nanoparticle is black, energy sedimentation completely under additional magnetic fields, and Zeta potential is+30 ± 5mV, three's Zeta potential result as shown in Figure 6, adds Al 3+the magnetic nano particle MNP1 that the Zeta potential of magnetic nano particle MNP2 and MNP3 of preparation is all prepared than traditional co-precipitation method is high, illustrates that aluminum ions adding adds coprecipitation generation Fe 3o 4zeta potential.
PH 5.0 cushioning liquid carries out as follows to the contrast of sedimentation situation after MNP1, MNP2 and MNP3 effect.
(1) acetic acid-ammonium acetate buffer of 0.1mol/L pH 5.0 is prepared
Get ammonium acetate 0.385g, the about 80ml that adds water adds glacial acetic acid 0.285ml after dissolving, and regulates pH to 5.0, be settled to 100ml, obtain final product by the NaOH solution of 3mol/L.
(2) get 500 μ L respectively and be about magnetic nano particle MNP1, MNP2 and MNP3 of 40mg/mL in 2mL centrifuge tube.
(3) Magneto separate, removes supernatant, adds the acetic acid-ammonium acetate buffer of 500 μ L 0.1mol/L pH 5.0 respectively, mixing.
(3) on blood mixer, mix 2 hours, Magneto separate, removes supernatant, and then clean once with 500 μ L ultra-pure waters, add 500 μ L ultra-pure waters, mixing, magnet compares three's sedimentation situation.
As shown in Figure 7, the magnetic nano particle MNP1 for preparing of traditional co-precipitation method can sedimentation completely, and supernatant is transparent for result, after add Al 3+the magnetic nano particle MNP3 major part of preparation can sedimentation, supernatant light brown, the magnetic nano particle MNP2 for preparing of the inventive method then can only sedimentation part, supernatant is black, illustrate that magnetic nano particle dispersiveness under pH 5.0 condition prepared by the inventive method improves, thus can not make its complete sedimentation, after add Al 3+the magnetic nano particle dispersiveness of preparation improves very little.
MNP1, MNP2 and MNP3 contrast (UV absorptiometry) to bovine serum albumin(BSA) BSA adsorbance is carried out as follows.
(1) get the centrifuge tube of 6 2mL, add magnetic nano particle MNP1 prepared by 3mg traditional co-precipitation method respectively, magnetic nano particle MNP2 prepared by the inventive method and after add Al 3+the magnetic nano particle MNP3 of preparation, often kind of magnetic nano particle 2 centrifuge tubes, clean 3 times with 1mL ultra-pure water respectively, outwell supernatant.
(2) take 210mg BSA, be dissolved in 7mL ultra-pure water, mixing obtains the BSA solution of 30mg/mL, is added by BSA solution in the centrifuge tube of magnetic nano particle place, each centrifuge tube 1mL, and then 6 centrifuge tubes are mixed 2h by mixing on blood mixer.
(3) all samples is carried out magnetic sedimentation, remove supernatant, then clean 1 time with 1mL ultra-pure water, remove supernatant, respectively add the disodium phosphate soln of 1mL 0.5mol/L, blood mixer mixes 2h and carries out desorption, the BSA that magnetic nano particle is adsorbed disintegrates down.
(4) magnetic sedimentation, get supernatant and carry out ultraviolet light absorption photometric detection, each sample surveys 3 times, and 280nm and 340nm locates the difference OD of absorption photometric value 280-340with the BSA concentration positive correlation in supernatant, can represent that different magnetic nano particle MNP1, MNP2 and MNP3 are to bovine serum albumin(BSA) BSA adsorbance.
As shown in Figure 8, the adsorbance of magnetic nano particle MNP2 to BSA prepared by the inventive method is the highest for result, and magnetic nano particle MNP1 prepared by traditional co-precipitation method and after add Al 3+the magnetic nano particle MNP3 of preparation is lower, adds Al after explanation 3+the absorption of magnetic nano particle to BSA can not be improved, first will add Al by the inventive method 3+prepare the magnetic nano particle that magnetic nano particle just can obtain high absorption.
Those skilled in the art will readily understand; the foregoing is only preferred embodiment of the present invention; not in order to limit the present invention, all any amendments done within the spirit and principles in the present invention, equivalent replacement and improvement etc., all should be included within protection scope of the present invention.

Claims (6)

1. a preparation method for superparamagnetic nano particle, is characterized in that, comprises the following steps:
(1) under starvation condition, by Fe 3+, Al 3+, Fe 2+be dispersed in water and form precursor solution, make Fe 3+concentration between 0.05mol/L to 0.5mol/L, Fe 3+and Fe 2+mol ratio be 2:1, Fe 3+and Al 3+mol ratio between 15:1 to 4:1;
(2) under starvation condition, by the precursor solution Keep agitation obtained in step (1), holding temperature 30 DEG C to 60 DEG C, and slowly dropping alkali lye regulates solution ph to be more than or equal to 9, forms suspension;
(3) under starvation condition, by the suspension Keep agitation obtained in step (2), and be warming up to 60 DEG C to 90 DEG C, reaction 0.5 is little of 2 hours, and cooling is precipitation and separation also, obtains described magnetic nanoparticle after cleaning.
2. the preparation method of magnetic nanoparticle as claimed in claim 1, is characterized in that, in step (2), mixing speed is 800rpm to 2000rpm, and the rate of addition of alkali lye is 0.5mL/min to 4mL/min.
3. the preparation method of magnetic nanoparticle as claimed in claim 1, is characterized in that, step (2) described alkali lye is the sodium carbonate liquor of pH >=9.5, ammonia spirit or sodium hydroxide solution.
4. the preparation method of magnetic nanoparticle as claimed in claim 1, it is characterized in that, step (2) and step (3) described stirring are mechanical agitation and/or ultrasonic vibration.
5. as the magnetic nanoparticle that Claims 1-4 any one preparation method prepares, it is characterized in that, described magnetic nanoparticle has superparamagnetism.
6. magnetic nanoparticle as claimed in claim 5, is characterized in that, the Zeta potential of described magnetic nanoparticle is between+25mV to+41mV.
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