CN104985193A - Alloy nano-particle preparation method based on protein reduction method - Google Patents
Alloy nano-particle preparation method based on protein reduction method Download PDFInfo
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Abstract
The invention belongs to an alloy nano-particle preparation method based on a protein reduction method. Firstly, 3-50mg ML<-1> of purified protein solution is prepared; 10-30 mM of precious metal salt solution formed by one of silver, gold, platinum or palladium or mixing any two of the silver, the gold, the platinum or the palladium is prepared; 20-30 mM of metal salt solution of copper, nickel, ferrous iron or aluminum is prepared; the purified protein solution, the precious metal salt solution and the metal salt solution are mixed to prepare a nano alloy precursor solution, the pH of the nano alloy precursor solution is adjusted to an alkaline range through a sodium hydroxide solution, the precursor solution is evenly mixed and then high-purity nitrogen is introduced for deoxygenization, then the mixture is placed in water bath to be heated, and nano alloy quickly reduced by protein can be obtained. Time for preparing precious metal nano-particles reduced by the protein is shortened from 1-5 h to 5-30 min, nano alloy with properties of target nano-particles can be quickly and effectively synthesized, the synthesis rate is greatly increased, and commercial application is facilitated.
Description
Technical field
The invention belongs to the preparation field of metal nanoparticle material, is a kind of alloy nanoparticle preparation method based on albumen reducing process.
Background technology
Binary or multicomponent alloy nano particle are with a wide range of applications in catalysis, optics, bio-sensing, drug delivery etc. with the physicochemical property of its excellence.Alloy not only can have character specific to single metal, also may produce synergy, so various novel nano alloying pellet combination emerges in an endless stream in recent years.
The synthesis of nano particle needs reducing agent and stabilizing agent.May bring negative effect to environment owing to adding chemical reducing agent, not meet the developing direction of Green Chemistry, therefore increasing green syt method is arisen at the historic moment.This comprises and utilizes various living resources, as: protein, DNA, plant extraction liquid, microorganism etc.Wherein, protein relies on the reproducibility amino acid residue of self can be directly used in the reduction of nano particle, is a kind of green gentle reduction means.But it expends time in longer, so still need developing new approaches to promote reaction rate and the productive rate of albumen direct-reduction nano particle.Also there is report about some novel preparation means, promoting that as utilized microwave method nano particle generates; Bubble is blasted in solution and form gas-liquid interface in order to synthesis of nano alloy etc.But the ion utilizing copper, nickel, ferrous iron, aluminium etc. easily to be reduced by albumen forms zeroth order nano-cluster, the noble metal such as Accelerating reduction silver, gold, platinum, palladium also synthesizes the alloy even method of ternary alloy three-partalloy and rarely has report.The alloy formed not only has spectral quality and the reducing property of former metal target nano particle, and the collaborative character of acquisition two kinds of nano metals of simultaneously also having an opportunity, application prospect is very wide.
Summary of the invention
The object of the invention is for the problems referred to above, develop a kind of method preparing alloy nanoparticle based on protein fast restore, under alkaline aqueous environment, prepare the alloy nanoparticle that diameter is 5-13nm.
Principle of the present invention is: based on copper, nickel, ferrous, aluminum ions standard electrode EMF is less than silver, gold, platinum, palladium ion, in order to improve the noble silver of the more difficult reduction of protein, gold, platinum, the generating rate of Pd nano particle and productive rate, utilize copper, nickel, ferrous, aluminum salt solution, use protein that they are reduced into copper, nickel, iron, aluminium nanocluster is as nucleation material, with the silver increased, gold, platinum, the rate of reduction of palladium ion and productive rate, synthesized alloy nanoparticle possesses the character of target noble metal nano particles, achieve the quick preparation of albumen direct-reduction alloy nanoparticle.
The alloy preparation method of a kind of albumen direct-reduction of the present invention comprises following concrete steps:
(1) purifying protein solution 3-50mg mL is prepared
-1;
(2) precious metal salt solution 10-30mM that are a kind of in silver, gold, platinum or palladium or wherein any two kinds of mixing are prepared;
(3) prepare the metal salt solution 20-30mM of copper, nickel, ferrous iron or aluminium, the standard redox potential of these metal needs noble metal middle than step (2) is low;
(4) (1) (2) (3) solution mixing system is become Nanoalloy precursor solution, regulate pH to alkaline range with sodium hydroxide solution, by high pure nitrogen deoxygenation logical after precursor solution Homogeneous phase mixing, then put into water-bath heating and can obtain by the Nanoalloy of albumen fast restore.
Record the surface plasma resonance spectrum peak of solution by ultraviolet specrophotometer or can monitor Product formation situation by transmission electron microscope, the provable synthesis of nano particle of the simple spike formed is homogeneous alloy.
In step (1), described protein solution is the proteins and peptides of the amino acid residue possessing reducing power containing pH within the scope of 8.0-13.0, in synthesis, do not limit it keep space structure, even the amyloid fiber body of these albumen is also all applicable.The amino acid wherein with reproducibility is tryptophan, tyrosine, lysine, arginine, histidine, cysteine, glutamine, choose high-quality cow's milk globulin, lysozyme, insulin, bovine serum albumin(BSA), and amyloid fiber formed by these albumen.What wherein mainly play reduction in a part cow's milk globulin is 2 trp residues, 4 tyrosine residues, 15 lysine residues; In a part lysozyme, a main reduction is 6 trp residues, 3 tyrosine residues, 6 lysine residues, 11 arginine residues; In a part insulin, a main reduction is 8 tyrosine residues, 2 lysine residues, 5 cysteine residues, 4 histidine residues; In a part bovine serum albumin(BSA), a main reduction is 20 tyrosine residues, 6 lysine residues, 6 glutamine residue.
In step (1), described protein solution compound method is: with ultra-pure water preparation 3-50mg mL
-1protein solution, regulates pH to 8.0-13.0, stablizes by its centrifuging and taking supernatant liquor after one hour, and by foreign ion in ultracentrifugation or the former albumen of dialysis removing.Final protein concentration is determined by ultraviolet specrophotometer.
In step (2), described precious metal salt solution is water-soluble monovalence silver salt, trivalent gold salt, tetravalence platinum salt, divalent palladium salt or both mixing arbitrarily among them; Preferred silver nitrate, gold chloride, chloroplatinic acid, palladium bichloride.
In step (3), described metal salt solution is water-soluble mantoquita, nickel salt, ferrous salt and aluminium salt; Preferred copper chloride, copper nitrate, copper sulphate, nickel chloride, frerrous chloride, aluminium chloride.
Described in step (1), protein solution and subject alloy product there is no special corresponding relation.
In step (4) described precursor solution, described precursor solution mixed volume ratio is purifying protein solution: precious metal salt solution: metal salt solution is 1:(0.2-18): (8-80).
The ultimate density of albumen is preferably 0.3-1mg mL
-1, metal ultimate density is preferably 500-1800 μM.
In step (4), the reaction temperature of described reaction is preferably 60-90 DEG C, and pH is preferably 8.0-13.0, exceed this pH scope, the amino acid residue playing reduction in albumen can be influenced, thus reduce nano particle productive rate, and described order by merging there is no particular/special requirement.
Compared with existing synthetic method, the invention has the advantages that:
(1) preparation time of albumen direct-reduction noble metal nano particles is reduced to 5-30min by 1-5h by the present invention, can fast and effeciently synthesize the Nanoalloy possessing target nanoparticle character, substantially increase synthesis rate;
(2) the invention provides a kind of synthesis utilizing the metallic copper of low electrode electromotive force, nickel, iron, aluminium to promote noble silver, gold, platinum, palladium nano-particles also final new approaches forming alloy, wherein protein is reducing agent and the stabilizing agent of nano particle;
(3) the present invention's protein used does not require the maintenance of space structure, and the protein of unfolding even amyloid filaments is also applicable to this synthetic method.
(4) this method preparation process is aqueous phase solution, organic solvent-free, utilizes protein for reducing agent, environmental protection, easy and simple to handle, and reduces the cost of synthesis noble metal nano particles;
(5) this method is except step 4 needs to carry out under the condition of 60-90 DEG C, and other all at room temperature carries out, and experiment condition is gentle;
(6) alloy nanoparticle prepared by this method is reproducible, is convenient to industrial production.
Accompanying drawing explanation
Fig. 1 for Kufil nano particle synthesized after adding copper sulphate described in the embodiment of the present invention 1 heat 5min in 90 DEG C of water-baths with do not add copper ion situation under in 90 DEG C of water-baths, heat 5min silver nano-grain Spectral Extinction contrast.After proof adds copper sulphate, reaction rate and productive rate significantly improve.
Fig. 2 is copper silver nanoparticle alloying pellet@cow's milk immunoglobulin complex high power transmission electron microscope image described in the embodiment of the present invention 1, and wherein (111) crystal face of the corresponding yellow gold of interplanar distance, demonstrates the synthesis of yellow gold.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described in further detail.Should be understood that this is only in order to demonstrate the invention, not limit the scope of the invention by any way.
Embodiment 1: the preparation of cow's milk globulin reduction copper-silver alloy nano particle, concrete operation step is as follows:
(1) prepare the cow's milk globulin solution of purifying: take a certain amount of cow's milk globulin powder and be dissolved in ultra-pure water, regulate pH to 11.0, stablize by its centrifuging and taking supernatant liquor after one hour, and by foreign ion in ultracentrifugation or the former albumen of dialysis removing.Finally, by ultraviolet specrophotometer, the cow's milk globulin solution concentration after purifying is defined as 4mg mL
-1.
(2) noble metal nano silver selects silver nitrate as presoma, preparation 10mM liquor argenti nitratis ophthalmicus.
(3) lower than silver-colored oxidation-reduction electrode electromotive force slaine selects copper sulphate as presoma, preparation 20mM copper-bath.
(4) alloy nano copper-Yin cow's milk immunoglobulin complex is prepared: by 500 μ L cow's milk globulin solutions, 180 μ L copper-baths, 300 μ L liquor argenti nitratis ophthalmicuses and 4mL deionized water are together placed in vial.Regulate pH value of solution to be 12.0, after magnetic agitation mixing, lead to high pure nitrogen deoxygenation to precursor solution.After mixing, in precursor solution, the concentration of cow's milk globulin is 0.4mg mL
-1.Then bottle is put into 90 DEG C of water-baths to heat, 15min can obtain the copper-silver alloy nano particle reduced by cow's milk globulin.
Do not add copper ion compared under same condition, heating 5min Kufil nanometer and silver nano-grain situation of synthesizing contrast as shown in Figure 1, show to add in the copper sulphate short time to reach very high yield.Its single surface plasma resonance spectrum peak position is at 410nm, simultaneously copper-silver nanometer particle high power transmission electron microscope image as shown in Figure 2, wherein (111) crystal face of the corresponding yellow gold of interplanar distance, proves to have synthesized homogeneous yellow gold nano particle.
Embodiment 2: the preparation of cow's milk globulin also native copper-Yin-Jin ternary alloy nano particle, concrete operation step is as follows:
(1) the cow's milk globulin solution of purifying is prepared: as in embodiment 1 as described in step (1), the cow's milk globulin after purifying is defined as 4mg mL
-1.
(2) noble metal nano gold and silver selects gold chloride and silver nitrate as presoma, preparation 10mM liquor argenti nitratis ophthalmicus, 20mM chlorauric acid solution.
(3) lower than the oxidation-reduction electrode electromotive force of gold, silver slaine selects copper chloride as presoma, preparation 20mM copper-bath.
(4) prepare alloy nano copper-Yin-Jin cow's milk immunoglobulin complex: by 500 μ L cow's milk globulin solutions, 180 μ L copper-baths, 240 μ L liquor argenti nitratis ophthalmicuses, 100 μ L chlorauric acid solutions, and 4mL deionized water is together placed in vial.Regulate pH value of solution to be 10, after magnetic agitation mixing, lead to high pure nitrogen deoxygenation to precursor solution.After mixing, in presoma, the concentration of cow's milk globulin is 0.4mg mL
-1.Then bottle is put into 80 DEG C of water-baths and heat the copper-Yin-Jin ternary alloy nano particle that 30min can obtain being reduced by cow's milk globulin.
Its single surface plasma resonance spectrum peak position, at 450nm, proves to successfully synthesize homogeneous copper-Yin-billon nano particle.
Embodiment 3: the preparation of cow's milk globulin amyloid fiber body reduction copper-silver alloy nano particle, concrete operation step is as follows:
(1) cow's milk globulin amyloid fiber body is prepared: as in embodiment 1, preparation concentration as described in step (1) is 50mg mL
-1purifying cow's milk globulin solution, its pH is adjusted to 12 and is placed on 20h in 90 DEG C of water-baths, obtain cow's milk globulin amyloid fiber solution.
(2) noble metal nano silver selects silver nitrate as presoma, preparation 30mM liquor argenti nitratis ophthalmicus.
(3) lower than silver-colored oxidation-reduction electrode electromotive force slaine selects copper nitrate as presoma, preparation 20mM copper-bath.
(4) alloy nano copper-Yin cow's milk globulin amyloid fiber nanocrystal composition is prepared: by 50 μ L cow's milk globulin amyloid fiber liquid solutions, 180 μ L copper-baths, 90 μ L liquor argenti nitratis ophthalmicuses, and 4mL deionized water is together placed in vial.Regulate pH value of solution to be 12, after magnetic agitation mixing, lead to high pure nitrogen deoxygenation to precursor solution.After mixing, in presoma, the concentration of cow's milk globulin amyloid fiber body is 0.4mgmL
-1.Then bottle is put into 80 DEG C of water-baths heat 15min can obtain by cow's milk globulin amyloid fiber body reduce copper-silver alloy nano particle.Its single surface plasma resonance spectrum peak position, at 410nm, proves to have synthesized homogeneous yellow gold nano particle.
Embodiment 4: the preparation of lysozyme reduction alloy nano nickel-palladium particle, concrete operation step is as follows:
(1) prepare the lysozyme soln of purifying: take a certain amount of lysozyme powder and be dissolved in ultra-pure water, regulate pH to 13, stablize by its centrifuging and taking supernatant liquor after one hour, and remove foreign ion in former albumen by ultracentrifugation.Finally, by ultraviolet specrophotometer, the lysozyme concentration after purifying is defined as 5mg mL
-1.
(2) noble metal nano palladium selects palladium bichloride as presoma, preparation 10mM palladium chloride solution.
(3) lower than palladium oxidation-reduction electrode electromotive force slaine selects nickel chloride as presoma, 20mM nickel chloride solution.
(4) prepare alloy nano nickel-palladium@lysozyme compound: by 500 μ L lysozyme solns, 100 μ L nickel chloride solutions, 240 μ L palladium chloride solutions, and 4mL deionized water is together placed in vial.Regulate pH value of solution to be 13, after magnetic agitation mixing, lead to high pure nitrogen deoxygenation to precursor solution.After mixing, in presoma, the concentration of lysozyme is 0.5mg mL
-1.Then bottle is put into 80 DEG C of water-baths and heat alloy nano nickel-palladium particle that 15min can obtain being reduced by lysozyme.
Embodiment 5: the preparation of bovine serum albumin(BSA) reduction alloy nano iron-platinum grain, concrete operation step is as follows:
(1) prepare the bovine serum albumin solution of purifying: take a certain amount of bovine serum albumin(BSA) powder and be dissolved in ultra-pure water, regulate pH to 12, stablize by its centrifuging and taking supernatant liquor after one hour, and remove foreign ion in former albumen by ultracentrifugation.Finally, by ultraviolet specrophotometer, the bovine serum albumin(BSA) concentration after purifying is defined as 3mg mL
-1.
(2) noble metal nano silver selects chloroplatinic acid as presoma, 10mM liquor argenti nitratis ophthalmicus.
(3) lower than platinum oxidation reducing electrode electromotive force slaine selects frerrous chloride as presoma, preparation 20mM solution of ferrous chloride.
(4) prepare alloy nano iron-Yin bovine serum albumin(BSA) compound: by 500 μ L bovine serum albumin solutions, 100 μ L solution of ferrous chloride, 200 μ L liquor argenti nitratis ophthalmicuses, and 4mL deionized water is together placed in vial.Regulate pH value of solution to be 12, after magnetic agitation mixing, lead to high pure nitrogen deoxygenation to precursor solution.After mixing, in presoma, the concentration of bovine serum albumin(BSA) is 0.3mg mL
-1.Then bottle is put into 90 DEG C of water-baths and heat alloy nano iron-Argent grain that 15min can obtain being reduced by bovine serum albumin(BSA).
Embodiment 6: the preparation of insulin reduced iron-billon nano particle, concrete operation step is as follows:
(1) prepare the insulin solutions of purifying: take a certain amount of insulin powder and be dissolved in ultra-pure water, regulate pH to 9, stablize by its centrifuging and taking supernatant liquor after one hour, and remove foreign ion in former albumen by ultracentrifugation.Finally, by ultraviolet specrophotometer, the insulin concentration after purifying is defined as 10mg mL
-1.
(2) noble metal nano gold selects gold chloride as presoma, preparation 20mM chlorauric acid solution.
(3) lower than golden oxidation-reduction electrode electromotive force slaine selects frerrous chloride as presoma, preparation 20mM solution of ferrous chloride.
(4) prepare alloy nano iron-Jin insulin complex substance: by 500 μ L insulin solutions, 100 μ L solution of ferrous chloride, 100 μ L chlorauric acid solutions, and 4mL deionized water is together placed in vial.Regulate pH value of solution to be 13, after magnetic agitation mixing, lead to high pure nitrogen deoxygenation to precursor solution.After mixing, in presoma, the concentration of insulin is 1mg mL
-1.Then bottle is put into 90 DEG C of water-baths and heat iron-billon nano particle that 10min can obtain being reduced by insulin.
Embodiment 7: the preparation of insulin amyloid fiber body also primary aluminum-silver alloy nano particle, concrete operation step is as follows:
(1) insulin amyloid fiber liquid solution is prepared: preparing concentration is as described in example 6 above 6mg mL
-1purified insulin solution, regulate pH to 2, adding NaCl to final concentration is 100mM, is placed in 60 DEG C of water-baths and heats 5h, obtain insulin amyloid fiber liquid solution.
(2) noble metal nano silver selects silver nitrate as presoma, preparation 10mM liquor argenti nitratis ophthalmicus.
(3) lower than silver-colored oxidation-reduction electrode electromotive force slaine selects aluminium chloride as presoma, 30mM liquor alumini chloridi.
(4) prepare alloy nano aluminium-Yin insulin amyloid fiber: by 500 μ L insulin corpus fibrosum solution, 100 μ L liquor alumini chloridis, 100 μ L liquor argenti nitratis ophthalmicuses, and 4mL deionized water is together placed in vial.Regulate pH value of solution to be 8, after magnetic agitation mixing, lead to high pure nitrogen deoxygenation to precursor solution.After mixing, in presoma, the concentration of insulin amyloid fiber body is 0.6mg mL
-1.Then bottle is put into 60 DEG C of water-baths and heat aluminium-silver alloy nano particle that 30min can obtain being reduced by insulin amyloid fiber.
A kind of alloy nanoparticle preparation method based on albumen reducing process that the present invention discloses and proposes, those skilled in the art are by using for reference present disclosure, the links such as suitable feed change and process route realize, although method of the present invention is described by preferred embodiment, person skilled obviously can change Method and Technology route as herein described or reconfigure not departing from content of the present invention, spirit and scope, realizes final technology of preparing.Special needs to be pointed out is, all similar replacements and change apparent to those skilled in the art, they are deemed to be included in spirit of the present invention, scope and content.
Claims (8)
1., based on an alloy nanoparticle preparation method for albumen reducing process, it is characterized in that comprising following process:
(1) purifying protein solution 3-50mg mL is prepared
-1;
(2) preparation is silver-colored, gold, platinum or palladium are a kind of or the precious metal salt solution 10-30mM of any two kinds of mixing;
(3) prepare a kind of metal salt solution 20-30mM in copper, nickel, ferrous iron, aluminium, the standard redox potential of the metal needs noble metal middle than step (2) of this step is low;
(4) (1) (2) (3) solution mixing system is become Nanoalloy precursor solution, regulate pH to alkaline range with sodium hydroxide solution, then pass into pure nitrogen gas deoxygenation, then put into the Nanoalloy that water-bath heating obtains being reduced by albumen.
2. the method for claim 1, is characterized in that: protein solution described in step (1) is cow's milk globulin, amyloid fiber formed by lysozyme, insulin, bovine serum albumin(BSA) and these albumen.
3. the method for claim 1, is characterized in that: be selected from water-soluble monovalence silver salt, trivalent gold salt, tetravalence platinum salt or divalent palladium salt for the precious metal salt solution described in step (2).
4. method as claimed in claim 3, is characterized in that precious metal salt solution is silver nitrate, gold chloride, chloroplatinic acid or palladium bichloride.
5. the method for claim 1, is characterized in that: the metal ion described in step (3) is selected from water-soluble mantoquita, nickel salt, ferrous salt or aluminium salt.
6. method as claimed in claim 5, is characterized in that described metal ion is selected from copper chloride, copper nitrate, copper sulphate, nickel chloride, frerrous chloride or aluminium chloride.
7. the method for claim 1, is characterized in that: the precursor solution mixed volume ratio described in step (4) is purifying protein solution: precious metal salt solution: metal salt solution is 1:(0.2-18): (8-80).
8. the method for claim 1, is characterized in that: in step (4), reaction system pH is 8.0-13.0, and reaction temperature is 60-90 DEG C.
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