CN103170641A - Method of preparing platinum nano particles by using cotton bollworm karyotype polyhedrin - Google Patents
Method of preparing platinum nano particles by using cotton bollworm karyotype polyhedrin Download PDFInfo
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Abstract
Provided is a method of preparing platinum nano particles by using cotton bollworm karyotype polyhedrin. Inherent structural features of the cotton bollworm karyotype polyhedrin are used, a PtC14 solution is added to a purified cotton bollworm karyotype polyhedral body solution, and the precious metal platinum nano particles which are even in distribution and consistent in diameter of the particles are obtained on the surface of the cotton bollworm karyotype polyhedrin through hatch, ultrasonic wave assistance reduction treatment. Due to the fact that a cotton bollworm karyotype polyhedron carrier has natural specific structures of functional group repetitive distribution, the complex preparation technology of a conventional carrier is avoided, and therefore production cost is reduced. The sizes and distribution of the precious metal platinum nano particles are effectively controlled through control of the alkaline hydrolysis degree of the cotton bollworm karyotype polyhedron, and the method of preparing the platinum nano particles by using the cotton bollworm karyotype polyhedrin can be significantly applied to catalytic fuel cells or biosensing or the like.
Description
Technical field
The present invention relates to a kind of preparations of nanomaterials method, particularly prepare the method for noble metal platinum nano particle.
Background technology
Noble metal nanometer material is owing to having good stability, small-size effect, skin effect, optical effect and the good characteristics such as biocompatibility, have important using value in fields such as optics, sensor, catalysis and biological detection analyses, thereby more and more get more and more people's extensive concerning.Size, pattern and composite construction thereof that the Recent study focus concentrates on by controlling nano particle prepare regulatable novel nano particle.At present the preparation method of noble metal platinum nano particle is a lot, but will obtain structure, form, size adjustable, is evenly distributed, the noble metal platinum nano particle is still more difficult preferably for stability.
In recent years, people often adopt the material with carbon element of different shape or other templates to control growth and the distribution of small particle diameter noble metal platinum nano particle as carrier.For example, people (the ACS.Appl.Mate.Interfaces.2011 such as Ping Lin K μ o, 3,115-118) and the people (Chem.Mater.2006 such as Robert V H μ ll, 18,1780-1788) reported respectively the nano platinum particle that utilizes porous carbon nanotube and multi-walled carbon nano-tubes preparation to disperse, because the material with carbon element itself as load lacks the activated adoption group, needed to introduce through processing early stage the group that can be combined with noble metal in preparation process.These operations make complicated process of preparation, have improved production cost, and the distribution of resulting metallic has very large randomness.Dan Wen etc. be published in " physical chemistry C " (J.Phys.Chem.C.2009,113,13023-13028) reported TiO in the article
2The preparation of colloidal spheres supporting Pt nano particle, the product of preparation can be used for the biology sensor material.Yet, due to TiO
2Itself lacks adsorption group, is difficult to make noble metal nano particles to be attached to its surface, need to add aminopropyl trimethoxysilane to introduce NH
2-Connect TiO as intermediate
2With noble metal.
Summary of the invention
The object of the present invention is to provide and a kind ofly can effectively control by the alkaline hydrolysis degree of controlling albumen the distribution of noble metal platinum nano particle, preparation condition is gentle, productive rate is high, technique is simple, and the method directly adopts the bioprotein nanostructured of nature existence as the carrier of noble metal platinum nano particle, wide material sources, repeatability is high, a kind of method of utilizing bollworm caryogram polyhedrin to prepare nano platinum particle that production cost is low.
The present invention adopts natural bollworm caryogram polyhedrin to control the noble metal platinum nano particle of synthetic high degree of dispersion as carrier.At first the present invention carries out the purifying alkaline hydrolysis to bollworm caryogram polyhedral body, obtains the polyhedrin sheet, then adds PtCl
4Solution carries out common hatching, make precious metal ion be adsorbed onto on particular functional group's's (as mercapto functional group) that caryogram polyhedrin surface contains avtive spot, process by ultrasonic auxiliary coreduction after the hatching special time, just formed at protein surface and arranged in order, the precious metals pt nano particle of size uniform.
Technical scheme of the present invention specifically comprises the steps:
(1) the polyhedrosis purifying of bollworm caryogram and alkaline hydrolysis:
The former powder of 0.5～1.0g heliothis armigera nuclear polyhedrosis virus is dissolved in 60～100mL ultra-pure water its dissolving of ultrasonic promotion.The present invention's ultra-pure water used prepares by the ultra-pure water instrument.Get supernatant after the centrifugal 5～10min of 500～1200r/min, get precipitation after the centrifugal 20～30min of 3000～5000r/min, precipitation is resuspended with ultra-pure water.So low speed, centrifugal 3～5 times repeatedly at a high speed, till last ultracentrifugal supernatant achromaticity and clarification, namely obtain purer polyhedral body.The gained polyhedral body is resuspended with the cold ultra-pure water of 1.5～2.5mL, add 150～250 μ L alkali solution liquids to process 10～25min.Above-mentioned alkali solution liquid is 0.3mol/L sodium carbonate, 0.5mol/L sodium chloride and 0.03mol/L disodium ethylene diamine tetra-acetic acid solution.Then regulate centrifugal 2～3 times of pH to 7～8,5000～7000r/min with the glacial acetic acid of 0.5mol/L, get supernatant.At 4～10 ℃, ultracentrifugation 1～2h under 25000～30000r/min after abandoning supernatant precipitates gained spot resuspended with 1～2mL ultra-pure water afterwards.
(2) the noble metal platinum nano particle of preparation bollworm caryogram polyhedrin load:
Getting bollworm caryogram polyhedrin liquid that 100～300 μ L alkaline hydrolysis process, to add 100～300 μ L concentration be the PtCl of 1～10mM
4In solution, abundant mixing, in 20～30 ℃, shaking table hatching 15～25h under 130～170r/min.Then dropwise adding the concentration of the new preparation of 50～100 μ L is the NaBH of 5～10mM
4Solution, limit ultrasonic limit reduction.After reduction, in 20～30 ℃, under 130～170r/min, shaking table hatching 3～6h, namely obtain bollworm caryogram polyhedrin carried noble metal nano platinum particle.
Because bioprotein comprises various active groups, amino acid sequence arranging rule, biodegradable, bio-compatible, has and improve and strict molecular recognition function; both can be used as the noble metal platinum nano particle that protective agent or adjusting control agent prepare different size, pattern in the preparation of noble metal platinum nano particle, can be used as again the regularly arranged noble metal platinum nano particle of guiding agent.Architectural feature and the space confinement effect thereof of biomolecule self can carry out accuracy controlling to the synthetic of nano particle simultaneously, thereby obtain having the nano particle of desired dimensions size and arranging rule.The bollworm caryogram polyhedrin that the present invention adopts is that heliothis armigera nuclear polyhedrosis virus produces in the later period of infection great expression, and self forms the super large molecule protein crystal of compact structure.Bollworm caryogram polyhedrin is that the trimerical subunit structure by a kind of densification consists of, and connects by disulfide bond between tripolymer.The theoretical length of a repetitive of this protein structures is 10nm, and each tripolymer length is 5nm.This specific structure repeatability is arranged, and its outer surface is rich in a large amount of amino acid, and contained active function groups can adsorb many kinds of metal ions to its surface.And distinguishing feature of this albumen is exactly to regulate by regulating the polyhedrosis alkaline hydrolysis time alkaline hydrolysis degree of albumen, and then controls the distribution of noble metal platinum nano particle.
Described noble metal platinum nano particle has following architectural feature: bollworm caryogram polyhedrin tactical rule, individual layer albumen presents orderly fenestral fabric, and protein surface has specific avtive spot easily to be combined with metal ion.Gained noble metal platinum nano particle is evenly distributed on protein surface, and decentralization is high, and the size of particle, decentralization are consistent with bollworm caryogram polyhedrin.
This method has not only reduced the preparation technology of traditional nano platinum particle carrier complexity, and need not to introduce the external source active group, the active function groups that albumen itself contains has namely been controlled the size of nano platinum particle as adjusting control agent, control arranging and decentralization of nano platinum particle as guiding agent again.
The invention has the beneficial effects as follows: directly adopt natural bollworm caryogram polyhedrin as carrier, need not carry out any specially treated, through hatching altogether with material system, ultrasonic auxiliary coreduction is processed, and can obtain the noble metal platinum nano particle of bollworm caryogram polyhedrin load.Compare as the noble metal platinum nano particle of carrier regulation and control growth with existing material with carbon element and porous oxide, the noble metal platinum nano particle that the present invention obtains has that size is little, particle diameter is more evenly distributed, disperse advantages of higher.This preparation method's technique is simple, mild condition, and environment-friendly high-efficiency, and the carrier wide material sources are easy to get, and with low cost, be easy to realize large-scale production.
Description of drawings
The invention will be further elaborated below in conjunction with drawings and Examples.
Fig. 1 is the TEM figure of the bollworm caryogram polyhedrin that obtains of alkaline hydrolysis 15min;
Fig. 2 is the TEM figure of nano platinum particle of bollworm caryogram polyhedrin load of the alkaline hydrolysis 15min of preparation;
Fig. 3 is the TEM figure of the bollworm caryogram polyhedrin that obtains of alkaline hydrolysis 25min;
Fig. 4 is the TEM figure of nano platinum particle of bollworm caryogram polyhedrin load of the alkaline hydrolysis 25min of preparation.
The specific embodiment
Embodiment 1
The former powder of 0.5g heliothis armigera nuclear polyhedrosis virus is dissolved in the 60mL ultra-pure water its dissolving of ultrasonic promotion.The PL5124-PALL model ultra-pure water instrument preparation that ultra-pure water used is produced by Britain's High Wycombe city pure water system Co., Ltd of Vivendi.Get supernatant after the centrifugal 10min of 1000r/min, get precipitation after the centrifugal 30min of 4000r/min, precipitation is resuspended with ultra-pure water.So low speed, centrifugal 4 times repeatedly at a high speed, till last ultracentrifugal supernatant achromaticity and clarification, namely obtain purer polyhedral body.The gained polyhedral body is resuspended with the cold ultra-pure water of 2mL, add 200 μ l alkali solution liquids to process 15min.Then regulate pH to 7 with the glacial acetic acid of 0.5mol/L, get supernatant after the centrifugal 5min of 5000r/min, 6000r/min is centrifugal, and 5min gets supernatant, and the centrifugal 5min of 7000r/min gets supernatant.At 4 ℃, ultracentrifugation 1h under 28000r/min after abandoning supernatant precipitates gained spot resuspended with the 1.5mL ultra-pure water afterwards.Observe its pattern under transmission electron microscope and see Fig. 1.Fig. 1 is the TEM figure of the bollworm caryogram polyhedrin after embodiment 1 purifying and alkaline hydrolysis, as can be seen from the figure the bollworm caryogram polyhedrin of gained presents the fenestral fabric of individual layer, single lattice are that a tripolymer assumes diamond in shape, be approximately square, size is 5nm, a plurality of tripolymer repeated arrangement form the reticuloprotein sheet, and the size of albumen flakes is 20～80nm, and main concentrating is 40～50nm.On each tripolymer, active function groups is arranged, for the forming core of noble metal provides the site, simultaneously its spacing effect limits in space the particle diameter of noble metal platinum nano particle be 4～5nm.
Getting bollworm caryogram polyhedrin liquid that 200 μ L alkaline hydrolysis process, to add 100 μ L concentration be the PtCl of 5mM
4Solution, abundant mixing, in 25 ℃, shaking table hatching 24h under 140r/min.Then dropwise adding the concentration of the new preparation of 75 μ L is the NaBH of 5mM
4Solution, limit ultrasonic limit reduction.After reduction, in 25 ℃, under 140r/min, shaking table is hatched 6h, namely obtains the nano platinum particle of bollworm caryogram polyhedrin load.Observe its pattern under transmission electron microscope and see Fig. 2.Fig. 2 is the TEM figure of nano platinum particle of the bollworm caryogram polyhedral body load of embodiment 1 preparation, nano platinum particle that can gained from figure disperses uniformly, size and shape and caryogram polyhedrin are basically identical, and single nano particle diameter is 4～5nm, there is no agglomeration.
Embodiment 2
The former powder of 0.5g heliothis armigera nuclear polyhedrosis virus is dissolved in the 60mL ultra-pure water its dissolving of ultrasonic promotion.The PL5124-PALL model ultra-pure water instrument preparation that ultra-pure water used is produced by Britain's High Wycombe city pure water system Co., Ltd of Vivendi.Get supernatant after the centrifugal 10min of 1000r/min, get precipitation after the centrifugal 25min of 4000r/min, precipitation is resuspended with ultra-pure water.So low speed, centrifugal 4 times repeatedly at a high speed, till last ultracentrifugal supernatant achromaticity and clarification, namely obtain purer polyhedral body.The gained polyhedral body is resuspended with the cold ultra-pure water of 1.5mL, add 200 μ l alkali solution liquids to process 15min.Then regulate pH to 7 with the glacial acetic acid of 0.5mol/L, get supernatant after the centrifugal 5min of 5000r/min, 6000r/min is centrifugal, and 5min gets supernatant, and the centrifugal 5min of 7000r/min gets supernatant.At 4 ℃, ultracentrifugation 1.5h under 28000r/min after abandoning supernatant precipitates gained spot resuspended with the 1.5mL ultra-pure water afterwards.
Getting bollworm caryogram polyhedrin liquid that 300 μ L alkaline hydrolysis process, to add 100 μ L concentration be the PtCl of 5mM
4Solution, abundant mixing, in 25 ℃, shaking table hatching 24h under 140r/min.Then dropwise adding the concentration of the new preparation of 75 μ L is the NaBH4 solution of 5mM, limit ultrasonic limit reduction.After reduction, in 25 ℃, under 140r/min, shaking table is hatched 6h, namely obtains the nano platinum particle of bollworm caryogram polyhedrin load.
Embodiment 3
The former powder of 0.5g heliothis armigera nuclear polyhedrosis virus is dissolved in the 60mL ultra-pure water its dissolving of ultrasonic promotion.The PL5124-PALL model ultra-pure water instrument preparation that ultra-pure water used is produced by Britain's High Wycombe city pure water system Co., Ltd of Vivendi.Get supernatant after the centrifugal 10min of 500r/min, get precipitation after the centrifugal 20min of 4000r/min, precipitation is resuspended with ultra-pure water.So low speed, centrifugal 3 times repeatedly at a high speed, till last ultracentrifugal supernatant achromaticity and clarification, namely obtain purer polyhedral body.The gained polyhedral body is resuspended with the cold ultra-pure water of 2mL, add 200 μ l alkali solution liquids to process 25min.Then regulate pH to 7 with the glacial acetic acid of 0.5mol/L, get supernatant after the centrifugal 5min of 5000r/min, 6000r/min is centrifugal, and 5min gets supernatant, and the centrifugal 5min of 7000r/min gets supernatant.At 4 ℃, ultracentrifugation 1h under 28000r/min after abandoning supernatant precipitates gained spot resuspended with the 1.5mL ultra-pure water afterwards.Observe its pattern under transmission electron microscope and see Fig. 3.Fig. 3 is the TEM figure of the bollworm caryogram polyhedrin after embodiment 3 purifying and alkaline hydrolysis, and as can be seen from the figure the alkaline hydrolysis time lengthens, and the albumen flakes of the caryogram polyhedrin of gained is more in small, broken bits, and main the concentrating of size is 10～20nm.
Getting bollworm caryogram polyhedrin liquid that 200 μ L alkaline hydrolysis process, to add 100 μ L concentration be the PtCl of 5mM
4Solution, abundant mixing, in 25 ℃, shaking table hatching 20h under 140r/min.Then dropwise adding the concentration of the new preparation of 75 μ L is the NaBH of 5mM
4Solution, limit ultrasonic limit reduction.After reduction, in 25 ℃, under 140r/min, shaking table is hatched 6h, namely obtains the nano platinum particle of bollworm caryogram polyhedrin load.Observe its pattern under transmission electron microscope and see Fig. 4.Fig. 4 is the TEM figure of nano platinum particle of the bollworm caryogram polyhedral body load of embodiment 3 preparation, nano platinum particle decentralization that can gained from figure is high, size and shape and caryogram polyhedrin are basically identical, and single nano particle diameter is 4～5nm, there is no agglomeration.
Embodiment 4
The former powder of 0.5g heliothis armigera nuclear polyhedrosis virus is dissolved in the 60mL ultra-pure water its dissolving of ultrasonic promotion.The PL5124-PALL model ultra-pure water instrument preparation that ultra-pure water used is produced by Britain's High Wycombe city pure water system Co., Ltd of Vivendi.Get supernatant after the centrifugal 5min of 500r/min, get precipitation after the centrifugal 30min of 3000r/min, precipitation is resuspended with ultra-pure water.So low speed, centrifugal 5 times repeatedly at a high speed, till last ultracentrifugal supernatant achromaticity and clarification, namely obtain purer polyhedral body.The gained polyhedral body is resuspended with the cold ultra-pure water of 1.5mL, add 150 μ l alkali solution liquids to process 10min.Then regulate pH to 7 with the glacial acetic acid of 0.5mol/L, get supernatant after the centrifugal 5min of 5000r/min, 6000r/min is centrifugal, and 5min gets supernatant, and the centrifugal 5min of 7000r/min gets supernatant.At 4 ℃, ultracentrifugation 1h under 25000r/min after abandoning supernatant precipitates gained spot resuspended with the 1mL ultra-pure water afterwards.
Getting bollworm caryogram polyhedrin liquid that 100 μ L alkaline hydrolysis process, to add 300 μ L concentration be the PtCl of 1mM
4Solution, abundant mixing, in 20 ℃, shaking table hatching 25h under 130r/min.Then dropwise adding the concentration of the new preparation of 50 μ L is the NaBH of 5mM
4Solution, limit ultrasonic limit reduction.After reduction, in 20 ℃, under 130r/min, shaking table is hatched 6h, namely obtains the nano platinum particle of bollworm caryogram polyhedrin load.
Embodiment 5
The former powder of 1.0g heliothis armigera nuclear polyhedrosis virus is dissolved in the 100mL ultra-pure water its dissolving of ultrasonic promotion.The PL5124-PALL model ultra-pure water instrument preparation that ultra-pure water used is produced by Britain's High Wycombe city pure water system Co., Ltd of Vivendi.Get supernatant after the centrifugal 10min of 1200r/min, get precipitation after the centrifugal 20min of 5000r/min, precipitation is resuspended with ultra-pure water.So low speed, centrifugal 3 times repeatedly at a high speed, till last ultracentrifugal supernatant achromaticity and clarification, namely obtain purer polyhedral body.The gained polyhedral body is resuspended with the cold ultra-pure water of 2.5mL, add 250 μ l alkali solution liquids to process 25min.Then regulate pH to 8 with the glacial acetic acid of 0.5mol/L, get supernatant after the centrifugal 5min of 5000r/min, 6000r/min is centrifugal, and 5min gets supernatant, and the centrifugal 5min of 7000r/min gets supernatant.At 10 ℃, ultracentrifugation 2h under 30000r/min after abandoning supernatant precipitates gained spot resuspended with the 2mL ultra-pure water afterwards.
Getting bollworm caryogram polyhedrin liquid that 300 μ L alkaline hydrolysis process, to add 100 μ L concentration be the PtCl of 10mM
4Solution, abundant mixing, in 30 ℃, shaking table hatching 15h under 170r/min.Then dropwise adding the concentration of the new preparation of 100 μ L is the NaBH of 10mM
4Solution, limit ultrasonic limit reduction.After reduction, in 30 ℃, under 170r/min, shaking table is hatched 3h, namely obtains the nano platinum particle of bollworm caryogram polyhedrin load.
Claims (2)
1. method of utilizing bollworm caryogram polyhedrin to prepare nano platinum particle, it is characterized in that: described method comprises the steps:
(1) the polyhedrosis purifying of bollworm caryogram and alkaline hydrolysis:
The former powder of 0.5～1.0g heliothis armigera nuclear polyhedrosis virus is dissolved in 60～100mL ultra-pure water its dissolving of ultrasonic promotion.Get supernatant after the centrifugal 5～10min of 500～1200r/min, get precipitation after the centrifugal 20～30min of 3000～5000r/min, precipitation is resuspended with ultra-pure water.Low speed like this, centrifugal 3～5 times repeatedly at a high speed, till last ultracentrifugal supernatant achromaticity and clarification, namely obtain purer polyhedral body, the gained polyhedral body is resuspended with the cold ultra-pure water of 1.5～2.5mL, add 150～250 μ l alkali solution liquids to process 10～25min.Then regulate centrifugal 2～3 times of pH to 7～8,5000～7000r/min with the glacial acetic acid of 0.5mol/L, get supernatant, afterwards at 4～10 ℃, ultracentrifugation 1～2h under 25000～30000r/min after abandoning supernatant precipitates gained spot resuspended with 1～2mL ultra-pure water;
(2) the noble metal platinum nano particle of preparation bollworm caryogram polyhedrin load:
Getting bollworm caryogram polyhedrin liquid that 100～300 μ L alkaline hydrolysis process, to add 100～300 μ L concentration be the PtCl of 1～10mM
4In solution, abundant mixing, in 20～30 ℃, shaking table hatching 15～25h under 130～170r/min.Then dropwise adding the concentration of the new preparation of 50～100 μ L is the NaBH of 5～10mM
4Solution, limit ultrasonic limit reduction.After reduction, in 20～30 ℃, under 130～170r/min, shaking table hatching 3～6h, namely obtain bollworm caryogram polyhedrin carried noble metal nano platinum particle.
2. the preparation method of the noble metal nano particles of caryogram polyhedrin according to claim 1 load, it is characterized in that: the alkali solution liquid in described step (1) is 0.3mol/L sodium carbonate, 0.5mol/L sodium chloride and 0.03mol/L disodium ethylene diamine tetra-acetic acid solution.
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| CN104001931A (en) * | 2014-05-12 | 2014-08-27 | 燕山大学 | Method for preparing precious metal palladium nanorod by means of cotton bollworm baculovirus nucleocapsid |
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| CN104741618A (en) * | 2015-02-13 | 2015-07-01 | 燕山大学 | Method for preparing platinum nanowires through helicoverpa armigera nuclear polyhedron extractives |
| CN114769610A (en) * | 2022-04-02 | 2022-07-22 | 西北工业大学 | Method for preparing gold-palladium nano alloy by using protein assembly |
| CN114854789A (en) * | 2022-05-06 | 2022-08-05 | 苏州大学 | Preparation method of nanocrystal-coated CyHv-2 nucleic acid subunit vaccine |
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| CN104668579A (en) * | 2015-02-11 | 2015-06-03 | 燕山大学 | Method for preparing palladium nano wires by use of heliothis armigera nuclear polyhedrin |
| CN104741618A (en) * | 2015-02-13 | 2015-07-01 | 燕山大学 | Method for preparing platinum nanowires through helicoverpa armigera nuclear polyhedron extractives |
| CN114769610A (en) * | 2022-04-02 | 2022-07-22 | 西北工业大学 | Method for preparing gold-palladium nano alloy by using protein assembly |
| CN114769610B (en) * | 2022-04-02 | 2023-08-11 | 西北工业大学 | Method for preparing gold-palladium nano alloy by utilizing protein assembly |
| CN114854789A (en) * | 2022-05-06 | 2022-08-05 | 苏州大学 | Preparation method of nanocrystal-coated CyHv-2 nucleic acid subunit vaccine |
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