CN102515276B - Method for preparing manganese dioxide nanoparticles with bovine serum albumin as template - Google Patents
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Abstract
The invention provides a preparation method for controlling to synthesize manganese dioxide nanoparticles with bovine serum albumin as a biological template, which belongs to the technical field of manganese dioxide synthesis and nano materials. A process has the following steps that: a certain amount of bovine serum albumin is dissolved in phosphate buffer solution, manganese acetate solution is added in, the mixed solution is mixed and reacts for 2min, then sodium hydroxide is added in, the pH value is adjusted to be strong alkaline, the reaction is carried out for 6h to 8h at room temperature, and the manganese dioxide nanoparticles with the grain size of 4.8nm are obtained after 2 days of dialysis and high-speed centrifugal separation. The method has the advantages that: the preparation method is simple, the reaction conditions are mild, the cost is low, little reagent is required, the use is environmental-friendly, and the manganese dioxide nanoparticles with good dispersibility, good stability, good biocompatibility, smaller grain size, consistent dimension and uniform shape and appearance are obtained. We think the manganese dioxide nanoparticles with smaller and uniform grain size have huge potential application prospect in absorption, catalysis, bio-marking and other fields.
Description
Technical field
The present invention relates to a kind of preparation method of manganese dioxide nano particle, belong to the synthetic and technical field of nano material of Manganse Dioxide, specifically a kind of is the method for the synthetic manganese dioxide nano particle of biological template contral based on bovine serum albumin.
Background technology
Growing along with nanosecond science and technology, the quantum size effect that nano particle has, small-size effect, surface effects and macroscopical channel effect etc. arouse widespread concern.These special character make nano material gather around and have broad application prospects at aspects such as optics, electricity, magnetics, catalysis.In recent years, Manganse Dioxide has caused the extensive concern in each field as a kind of oxide compound with essential industry purposes.Therefore, nano-manganese dioxide with its unique structure, excellent performance and cheap cost at secondary cell, stealthy, magneticsubstance, catalyzer, sorbent material, ion-exchange, wet sensitive and gas sensor, communication, high frequency circuit, (V. Bharat even all is widely used at the aspects such as colorant of paint or coating, R. Boppana and F. Jiao
Chem. Commun., 2011,47,8973-8975; F. Y. Cheng, J. Chen, X. L. Gou and P. W. Shen,
Adv. Mater., 2005,17,2753-2756.).
Find after deliberation, the granular size of nanometer titanium dioxide manganese material, pattern and crystal formation are very large to its performance impact.Therefore, preparation method's tool of research nano material of manganese dioxide has very important significance.At present, existing bibliographical information synthesized various nanometer rod, nano wire, nano belt, nanotube and 3-D nano, structure manganese bioxide material (X. Wang and Y. Li,
Chem. Commun., 2002,7,764-765; R. Ma, Y. Bando, L. Zhang and T. Sasaki,
Adv. Mater., 2004,16,918-922; Z. Liu, R. Ma, Y. Ebina, K. Takada and T. Sasaki,
Chem. Mater., 2007,19,6504-6512; D. Zheng, S. Sun, W. Fan, H. Yu, C. Fan, G. Cao, Z. Yin and X. Song,
J. Phys. Chem. B, 2005,109,16439-16443.).On the whole, the method for synthetic nano material of manganese dioxide mainly comprises: via sol-gel template method (B. B. Lakshmi, C. J. Patrissi and C. R. Martin,
Chem. Mater., 1997,9,2544-2550.), thermal decomposition method (D. Zitoun, N. Pinna, N. Frolet and C. Belin,
J. Am. Chem. Soc., 2005,127,15034-15035.), circumfluence method (J. C. Villegas, L. J. Garces, S. Gomez, J. P. Duand and S. L. Suib,
Chem. Mater., 2005,17,1910-1918.), solid phase method (Y. S. Ding, X. F. Shen, S. Sithambaram, S. Gomez, R. Kumar, V. M. Crisostomo and S. L. Suib,
Chem. Mater., 2005,17,5382-5389.), electrochemical deposition method (M. S. Wu, J. T. Lee, Y. Y. Wang and C. C. Wan,
J. Phys. Chem. B, 2004,108,16331-16333.) and hydrothermal method (X. Wang and Y. Li,
Chem. Eur. J., 2003,9,300-306.) etc.Make a general survey of the whole bag of tricks of synthetic nano material of manganese dioxide, great majority need the condition of High Temperature High Pressure usually, complicated operation, and generally need the reagent used more, caused waste.Wayward due to speed of response, products therefrom often exceeds nanometer range, and the reunion that particle easily occurs causes dispersed bad.And in the application of nano material, catalytic field especially, the small size of homogeneous and polymolecularity are vital.Regrettably, at present, be less than the particle diameter of 5 nm and the preparation method of dispersed extraordinary manganese dioxide nano particle has no report for synthetic.Therefore, the nano material of manganese dioxide that how utilizes at normal temperatures and pressures easy operation to obtain small particle size be one highly significant and be rich in challenging research work.
In recent years, the standby nano material of biological template legal system has caused attention widely.Biological tissue and biomacromolecule that the biological template nano materials refers to have ad hoc structure are template, utilize biological self-assembly and space confinement effect thereof, by methods such as physics, chemistry according to design requirements to size, pattern, the structure of synthetic materials, arrange etc. and to control, thereby form new nano material or nanostructure (Ou Yangjianming
Chemical progress, 2005,17 (4), 749-756.).Wherein, protein molecule is owing to having complicated space structure and multiple different biological function, for the pattern of nano material play vital effect (Xue Zhonghui, Dang Yazheng, CHEN XINHUA, Guan Rongfeng,
Xuchang College's journal, 2008,27 (5), 107-111.).Therefore, Study on Protein regulation and control nanoparticle synthetic enjoys favor.Be subjected to the process of protein regulation synthesis oxide nanoparticle generally can be divided into ionic bonding, oxidation, nucleation, growth four-stage (M. B. Dickerson, K. H. Sandhage and R. R. Naik,
Chem. Rev., 2008,108,4935-4978.).In regulation and control nanoparticle nucleation process, amino-acid residue in prlmary structure of protein plays an important role to the identification of mineral ion, secondary protein structure can adapt to by the variation of conformation the molecular structure of mineral crystal, thereby reach purpose (the L. B. Gower of the oriented growth of the regulation and control nucleation of crystal and crystal face
Chem. Rev., 2008,108,4551-4627.).The protein of take is as the advantage of the method for biological template synthesizing nano-particle: the reaction conditions gentleness, do not need heating and pressurizing, and generally can react at normal temperatures; Specific, the good biocompatibility of appearance of nano material rule, uniform particle diameter, crystal formation of preparation, have the prospect of biological fields such as being applied to immune labeled, bio-imaging, pharmaceutical carrier.
At present, the biological template method was widely used in simple, green characteristic in synthesizing of the multiple inorganic materials such as noble metal nano particles, oxide compound, sulfide, mixture.But up to now, utilize the method for the synthetic manganese dioxide nano particle of biological template method to have no report.
Summary of the invention
The objective of the invention is to solve the defects such as experiment condition harshness in the synthetic Manganse Dioxide method of tradition, complicated operation, dispersed bad, easy reunion, particle diameter are larger, it is the simple method for preparing of the Manganse Dioxide of 4.8 nm for biological template contral synthesizes particle diameter that a kind of bovine serum albumin that utilizes is provided.The present invention utilizes self-assembly characteristics, the space confinement effect of biological template and improves and strict molecular recognition function, by physics, chemical action, the synthetic of nano material carried out to accuracy controlling, thereby obtain having the nano material of manganese dioxide of specific morphology and structure.
For achieving the above object, the present invention is by the following technical solutions:
The preparation method of bovine serum albumin regulation and control manganese dioxide nano particle: in the process of preparation, add the phosphate buffer solution of bovine serum albumin in 50 mL single port bottles.Then add the aqueous solution that manganese acetate the is made into source as mn ion, on magnetic stirring apparatus, at the uniform velocity stir 2 min, temperature of reaction is 20 ℃ of normal temperature.Then adding aqueous sodium hydroxide solution to regulate pH is strong basicity, from colourless yellowing, continues to stir 6-8 hour after the solution several minutes, and solution becomes stabilising system.Through dialysis 48 hours, then high speed centrifugation separated, and can obtain the nanoparticle Manganse Dioxide of uniform particle diameter.Be kept in 4 ℃ of conditions and store.
The present invention has following beneficial effect:
A) the prepared nano material Manganse Dioxide of the present invention has the particle diameter of less homogeneous, good dispersiveness and stability, and the not sedimentation of standing some months, do not produce agglomeration.
B) template of the present invention is the protein bovine serum albumin, nontoxic, a kind of biological template reagent of green, and, by the synthetic manganese dioxide nano particle good biocompatibility of this biological template method, can be applicable in the biological fields such as biomarker, immunoassay, bio-imaging analysis.
C) preparation method's mild condition of the present invention, cost low, simple to operate, can react at normal temperatures and pressures, not need to add in addition organic stabilizer/dispersion agent, be conducive to the widespread use of nanoparticle in biological field.With the method for the synthetic Manganse Dioxide of tradition, comparing, overcome that traditional building-up reactions is complicated, the time is long, need that reagent is many, power consumption, the first-class shortcoming of particle diameter inequality, is a kind of synthetic method of simple green.
D) because the present invention has versatility, not only utilize protein molecule can control synthetic other oxide compounds such as zinc oxide, ferric oxide, Cadmium oxide, plumbous oxide, cupric oxide etc. for template, and utilize different protein moleculars can synthesize the inorganic nano material of different-shape, particle diameter, crystal formation, character for template, be a kind of synthetic method of very effective control nanoparticle.
The accompanying drawing explanation
Fig. 1 a, Fig. 1 b and Fig. 1 c show respectively transmission electron microscope (TEM) photo, high-resolution-ration transmission electric-lens (HRTEM) photo and the selected area electron diffraction figure (SAED) of products therefrom.From a figure, finding out, the manganese dioxide nanocrystal that the bovine serum albumin of take makes as template is as spherical, and median size is 4.8 nm and dispersed fine.From data analysis b figure, show that spacing is 0.214 nm, with γ-MnO
2In hexagonal, the spacing of (3 0 1) crystal face matches.Spacing and the γ-MnO of two diffraction rings in c figure
2The spacing of (3 0 1) and (the 31 0) crystal face in the X-ray diffraction of hexagonal (XRD) standard card matches, and proves that products therefrom is γ-MnO
2.
Fig. 2 is the dynamic light scattering figure of product.As can be seen from the figure the median size of Manganse Dioxide is 4.8 nm, and the hydration particle diameter is 7.3 nm.Particle diameter distribution width is narrow, and the Manganse Dioxide uniform particle diameter that the method is synthetic is described.
Fig. 3 is electronic spectrum (a) and the photoelectron spectrum figure (b, c, d) of product.From a and b figure, finding out, the main component of product is carbon, nitrogen, oxygen, sulphur, manganese.From c figure, finding out, in conjunction with can 642.4 eV and 653.9 eV be respectively in Manganse Dioxide Mn 2p3/2 and Mn 2p1/2 in conjunction with can value.D figure is the differential matched curve of sample O1s.As can be seen from the figure, in this sample, there are three kinds of combinations in oxygen, is respectively Mn-O-Mn key, Mn-O-H key and H-O-H key, and further the indirect proof product is Manganse Dioxide.
Fig. 4 is the circular dichroism spectrogram of product.The a figure line means is the circular dichroism spectrogram of pure bovine serum albumin, occurs respectively that at 208 and 222 nm two negative feature acromion bands of a spectrum are characteristic peaks of protein α-helixstructure.What b figure meaned is the circular dichroism spectrum peak that bovine serum albumin is controlled synthetic Manganse Dioxide, the peak of finding 208 nm has moved to 204 nm places, illustrates that protein is in the process of the synthetic Manganse Dioxide of regulation and control, and variation has occurred secondary structure, α-helixstructure reduces gradually, and the beta sheet structure increases.Major cause is the sterically hindered α-helixstructure that is less than of the secondary structure of beta sheet, more is conducive to nanoparticle and enters in protein steric structure and react.So obtaining an important conclusion is that variation has occurred the secondary structure of bovine serum albumin in the process of controlling synthetic Manganse Dioxide.
Fig. 5 is the infrared spectrum of product.As can be seen from the figure, b figure is the infrared spectrum of pure bovine serum albumin, 3311,3071,1654,1540 cm
-1Be respectively-OH ,-the molecule stretching vibration peak of NH-, acid amides I, acid amides II.A figure is the infrared spectrum that bovine serum albumin is controlled synthetic manganese dioxide nano particle.With the contrast of pure bovine serum albumin, can find out-OH and-movement occurred in the stretching vibration peak of NH-.Explanation in controlling synthetic process, mn ion and-OH and-interaction occurred in NH-, thereby take protein has formed nano material of manganese dioxide as template.
Embodiment
Below by embodiment, the preparation method who is the synthetic manganese dioxide nano particle of biological template contral based on bovine serum albumin of the present invention is described further.The present embodiment is implemented take the present invention program under prerequisite, provided detailed embodiment and concrete operating process.
Embodiment 1:
In the process of preparation, 10 mL that add respectively 25 mg bovine serum albumins to be made into, 0.01 mol L
-1Phosphate buffer solution in 4 50 mL single port bottles, then add respectively 5 μ mol manganese acetates, Manganous chloride tetrahydrate, manganous sulfate or manganese nitrate solution, on magnetic stirring apparatus, at the uniform velocity stir 2 min, temperature of reaction is 20 ℃ of normal temperature.Then add 50 μ L, 1 mol L
-1Aqueous sodium hydroxide solution, continue to stir 6-8 hour, and solution becomes stabilising system.Through dialysis, with high speed centrifugation, separated in 48 hours, can obtain the nanoparticle Manganse Dioxide of uniform particle diameter.Be kept in 4 ℃ of conditions and store.
Embodiment 2:
In the process of preparation, 10 mL that add respectively 25 mg bovine serum albumins to be made into, 0.01 mol L
-1Phosphate buffer solution in 3 50 mL single port bottles, then add respectively 5 μ mol manganese acetate solution, on magnetic stirring apparatus, at the uniform velocity stir 2 min, temperature of reaction is 20 ℃ of normal temperature.Then add respectively 50 μ L, 1 mol L
-1Aqueous sodium hydroxide solution, potassium hydroxide aqueous solution or ammoniacal liquor, continue to stir 6-8 hour, and solution becomes stabilising system.Through dialysis, with high speed centrifugation, separated in 48 hours, can obtain the nanoparticle Manganse Dioxide of uniform particle diameter.Be kept in 4 ℃ of conditions and store.
Embodiment 3:
In the process of preparation, 10 mL that add respectively 25 mg bovine serum albumins to be made into, 0.01 mol L
-1Phosphate buffer solution in 11 50 mL single port bottles, then add 5 μ mol manganese acetate solution, on magnetic stirring apparatus, at the uniform velocity stir 2 min, temperature of reaction is 20 ℃ of normal temperature.Then the 1 mol L that adds respectively different amounts
-1Aqueous sodium hydroxide solution is regulated pH and is respectively 8.10,8.35,9.06,10.00,10.68,10.90,11.28,11.38,11.47,11.54,11.62, continues to stir 6-8 hour, and solution becomes stabilising system.Through dialysis, with high speed centrifugation, separated in 48 hours, can obtain the nanoparticle Manganse Dioxide of uniform particle diameter.Be kept in 4 ℃ of conditions and store.
Embodiment 4:
In the process of preparation, 10 mL that add respectively 25 mg bovine serum albumins to be made into, 0.01 mol L
-1Phosphate buffer solution in 10 50 mL single port bottles, then add respectively 5 μ mol manganese acetate solution, on magnetic stirring apparatus, at the uniform velocity stir 2 min, temperature of reaction is respectively 10,20,30,40,50,60,70,80,90,100 ℃.Then add 1 mol L
-1Aqueous sodium hydroxide solution 50 μ L.Continue to stir 6-8 hour, solution becomes stabilising system.Through dialysis, with high speed centrifugation, separated in 48 hours, can obtain the nanoparticle Manganse Dioxide of uniform particle diameter.Be kept in 4 ℃ of conditions and store.
Embodiment 5:
In the process of preparation, the concentration that adds 25 mg bovine serum albumins to be made into 10 mL is respectively 0.00001,0.0001,0.001,0.01,0.1., 1 mol L
-1Phosphate buffer solution in 6 50 mL single port bottles, then add respectively 5 μ mol manganese acetate solution, on magnetic stirring apparatus, at the uniform velocity stir 2 min, temperature of reaction is 20 ℃ of normal temperature.Then add 1 mol L
-1Aqueous sodium hydroxide solution 50 μ L, continue to stir 6-8 hour, and solution becomes stabilising system.Through dialysis, with high speed centrifugation, separated in 48 hours, can obtain the nanoparticle Manganse Dioxide of uniform particle diameter.Be kept in 4 ℃ of conditions and store.
Embodiment 6:
In the process of preparation, add the bovine serum albumin of different amounts to be respectively the 0.01 mol L that 5,25,50,100,125,150 mg are made into 10 mL
-1Phosphate buffer solution in 6 50 mL single port bottles, then add respectively 25 μ mol manganese acetate solution, on magnetic stirring apparatus, at the uniform velocity stir 2 min, temperature of reaction is 20 ℃ of normal temperature.Then add 1 mol L
-1Aqueous sodium hydroxide solution 250 μ L, continue to stir 6-8 hour, and solution becomes stabilising system.Through dialysis, with high speed centrifugation, separated in 48 hours, can obtain the nanoparticle Manganse Dioxide of uniform particle diameter.Be kept in 4 ℃ of conditions and store.
Claims (4)
1. one kind is the preparation method of the synthetic manganese dioxide nano particle of biological template contral based on bovine serum albumin, it is characterized in that, preparation process comprises:
Bovine serum albumin is fitted over to 0.01 mol L
-1In phosphoric acid buffer, be made into 2.5 mg mL
-1Solution, then be placed in 4 ℃ of Refrigerator stores; Get above-mentioned bovine serum albumen solution 10 mL in 50 mL single port bottles, add 50 μ L, 0.1 mol L
-1Bivalent manganese solution is as the mn ion source, room temperature magnetic agitation 2 min, and stirring is that the participation of dissolved oxygen is prerequisite in order to make the dissolved oxygen attend fully in building-up process; Add 50 μ L, 1 mol L
-1It is strong basicity that alkali lye is regulated pH, and the pH value is 11.62, reaction 6-8 hour under normal temperature and pressure, the yellow solution of the manganese dioxide nano particle that can obtain disperseing.
2. described according to right 1 is the preparation method of the synthetic manganese dioxide nano particle of biological template contral based on bovine serum albumin; it is characterized in that; the employing bovine serum albumin is biological template; rather than other biomolecules; do not need High Temperature High Pressure ventilation protection; directly normal-temperature reaction on magnetic stirring apparatus, prepare the manganese dioxide nano particle.
3. described according to right 1 is the preparation method of the synthetic manganese dioxide nano particle of biological template contral based on bovine serum albumin, it is characterized in that, the manganese dioxide nano particle molecular weight cut-off of packing into of preparation is in the dialysis tubing of 8000-14000, dialysis tubing is immersed in redistilled water, and dialysis is 2 days under agitation condition.
4. described according to right 1 is the preparation method of the synthetic manganese dioxide nano particle of biological template contral based on bovine serum albumin, it is characterized in that, the manganese dioxide nano particle after dialysis is by high speed centrifugation, and rotating speed is 18500 rpm, times 30 min, remove unreacted bovine serum albumin.
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| CN112691120B (en) * | 2019-10-23 | 2022-03-29 | 北京大学 | Application of bivalent manganese in preparation of immune enhancement medicine or anti-tumor medicine |
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