CN105108137B - A kind of preparation method of the nano particle of strong catalase activity - Google Patents
A kind of preparation method of the nano particle of strong catalase activity Download PDFInfo
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- CN105108137B CN105108137B CN201510616671.5A CN201510616671A CN105108137B CN 105108137 B CN105108137 B CN 105108137B CN 201510616671 A CN201510616671 A CN 201510616671A CN 105108137 B CN105108137 B CN 105108137B
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Abstract
The invention discloses a kind of preparation method of the nano particle of strong catalase activity, comprise the following steps：(1) by chlorauric acid solution boiling reflux with continuous stirring, while at the uniform velocity adding sodium citrate solution to form particle diameter 12~14nm gold nano grains to reduce gold chloride；(2) to the mixed solution that gold chloride and chloroplatinic acid are added in material obtained in step (1), it is then slowly added into ascorbic acid solution, with the 2~12h of speed stirring reaction of 100~800rpm at a temperature of 10~50 DEG C, so as to wrap up golden platinum bimetallic shell on above-mentioned gold nano grain, the nano particle of the strong catalase activity is obtained final product.Compared with conventional method, this method is simple, efficient, highly versatile for the present invention, and the nano particle of synthesis has strong and stabilization catalase activity.
Description
Technical field
The invention belongs to biologically active nanometer field of material technology, and in particular to a kind of nanometer of strong catalase activity
The preparation method of particle.
Background technology
Catalase is that generally existing can be catalytically decomposed the egg that hydrogen peroxide produces oxygen and water in a kind of organism
White enzyme.(George,P.Reaction between Catalase and Hydrogen Peroxide[J].Nature
1947,160,41-43；Lemberg,R.；Foulkes,E.C.Reaction between Catalase and Hydrogen
Peroxide [J] .Nature 1948,161,131-132) at present, catalase due to strong catalase activity
It is widely used in the analysis detection of various targets.(Zhu,Z.；Guan,Z.；Jia,S.；Lei,Z.；Lin,S.；Zhang,H.；
Ma,Y.；Tian,Z.-Q.；Yang,C.J.Au@Pt Nanoparticle Encapsulated Target-Responsive
Hydrogel with Volumetric Bar-Chart Chip Readout for Quantitative Point-of-
Care Testing[J].Angew.Chem.,Int.Ed.2014,53,12503-12507；Song,Y.；Zhang,Y.；
Bernard,P.E.；Reuben,J.M.；Ueno,N.T.；Arlinghaus,R.B.；Zu,Y.；Qin,L.Multiplexed
Volumetric Bar-Chart Chip for Point-of-Care Diagnostics[J].Nat.Commun.2012,3,
1-9；Song,Y.；Xia,X.；Wu,X.；Wang,P.；Qin,L.Integration of Platinum Nanoparticles
with a Volumetric Bar-Chart Chip for Biomarker Assays[J].Angew.Chem.,
Int.Ed.2014,53,12451-12455；Song,Y.；Wang,Y.；Qin,L.A Multistage Volumetric Bar
Chart Chip for Visualized Quantification of DNA[J].J.Am.Chem.Soc.2013,135,
But 16785-16788), catalase there is also some shortcomings：First, as a kind of albumen, catalase is in cell
Easily it is degraded in the complex system such as lysate or serum；Secondly, the secondary structure of catalase easily by extreme environment such as high temperature,
The influence of the factor such as strong acid and highly basic, makes catalytic efficiency that irreversible decline to occur；(Song,Y.；Wang,Y.；Qin,L.A
Multistage Volumetric Bar Chart Chip for Visualized Quantification of DNA[J]
.J.Am.Chem.Soc.2013,135,16785-16788；Melov,S.；Ravenscroft,J.；Malik,S.；Gill,
M.S.；Walker,D.W.；Clayton,P.E.；Wallace,D.C.；Malfroy,B.；Doctrow,S.R.；Lithgow,
G.J.Extension of Life-Span with Superoxide Dismutase/Catalase Mimetics[J]
.Science 2000,289,1567-1569) it is last, prepared using bioengineering, purifying and preserving catalase protects it
Hold higher degree and catalysis activity is relatively costly.Therefore, the synthetic method for developing strong catalase activity nano particle has weight
The meaning wanted.
The content of the invention
It is an object of the invention to overcome prior art defect, there is provided a kind of nano particle of strong catalase activity
Preparation method.
Concrete technical scheme of the invention is as follows：
A kind of preparation method of the nano particle of strong catalase activity, comprises the following steps：
(1) by chlorauric acid solution boiling reflux with continuous stirring, while at the uniform velocity adding sodium citrate solution to reduce chlorine
Auric acid forms particle diameter 12~14nm gold nano grains, and wherein chlorauric acid solution and the volume ratio of sodium citrate solution is 100:1~
Gold chloride containing 0.01~0.02wt% in 1.5, and chlorauric acid solution, contains 2.8~3.5wt%'s in sodium citrate solution
Sodium citrate；
(2) to the mixed solution that gold chloride and chloroplatinic acid are added in material obtained in step (1), it is then slowly added into anti-bad
Hematic acid solution, with the speed 2~12h of stirring reaction of 100~800rpm at a temperature of 10~50 DEG C, so that above-mentioned gold nano grain
The upper golden platinum bimetallic shell of parcel, obtains final product the nano particle of the strong catalase activity, wherein in the reactant of the step
In system, the concentration of chloroplatinic acid is 0.04~0.064mM, and the concentration of gold chloride is 0.2~3.2mM, the concentration of ascorbic acid for 1~
16mM, and the mass ratio of above-mentioned chloroplatinic acid and gold chloride is 10~40:100, the gross mass and Vitamin C of chloroplatinic acid and gold chloride
The mass ratio of acid is 13.4~53.5:100, the mass ratio of gold nano grain and chloroplatinic acid and gold chloride gross mass for 2.7~
11.2:100。
In a preferred embodiment of the invention, the step (1) is：Chlorauric acid solution is boiled with continuous stirring
Boiling backflow, while at the uniform velocity adding sodium citrate solution to form particle diameter 13nm gold nano grains, wherein gold chloride to reduce gold chloride
Solution is 100 with the volume ratio of sodium citrate solution:Gold chloride containing 0.01wt%, sodium citrate in 1, and chlorauric acid solution
Sodium citrate containing 3wt% in solution.
It is further preferred that the temperature in the step (2) is 20~40 DEG C.
It is further preferred that the mixing speed in the step (2) is 300~500rpm.
It is further preferred that the reaction time in the step (2) is 5~9h.
It is further preferred that the concentration of chloroplatinic acid is 0.08~0.32mM in the step (2).
It is further preferred that the concentration of gold chloride is 0.4~1.6mM in the step (2).
It is further preferred that the concentration of ascorbic acid is 2~8mM in the step (2).
The beneficial effects of the invention are as follows：
1st, preparation method of the invention is catalyzed from the very strong material of this catalase activity of platinum as nano particle
Activated centre, making the nano particle of synthesis has very strong catalase activity；
2nd, preparation method of the invention wraps up the bimetallic shell of gold and platinum using gold and the synergy of platinum on gold seeds
Layer, further increases the catalase activity of nano particle；
3rd, the characteristics of preparation method of the invention interacts strong using functional groups such as gold and amino, sulfydryl and carboxyls, makes
The nano particle of synthesis is easy to modified biological macromolecular.
4th, compared with conventional method, this method is simple, efficient, highly versatile, the nanometer of synthesis for preparation method of the invention
Grain has strong and stabilization catalase activity, is the nano particle with catalase activity in bioanalysis and biology
Application in medical science provides new platform.
Brief description of the drawings
Fig. 1 is the composition principle figure of the nano particle of strong catalase activity of the invention.
(A) is the gold nano grain and obtained Au@AuPtNPs of step (1) preparation in the embodiment of the present invention 1 in Fig. 2
Uv-visible absorption spectra phenogram；(B) the X-ray energy spectrum phenogram of the Au@AuPtNPs prepared for the embodiment of the present invention 1.
The transmission electron microscope picture of 13nm gold nano grains prepared by the step of (A) is the embodiment of the present invention 1 in Fig. 3 (1)；(B)
It is the transmission electron microscope picture of the Au@AuPtNPs of the preparation of the embodiment of the present invention 1；(C) for the embodiment of the present invention 1 preparation Au@
The high-resolution-ration transmission electric-lens figure of AuPtNPs；(D) for the embodiment of the present invention 1 preparation Au@AuPtNPs SEAD
Figure.
Fig. 4 is the gold element (A) and the X-ray light of platinum element (B) on Au@AuPtNPs obtained in embodiments of the invention
Electron spectrum phenogram.
Fig. 5 is the experimental condition optimization procedure chart of the embodiment of the present invention 1, wherein：(A) various concentrations gold chloride and chloroplatinic acid；
(B) various concentrations ascorbic acid；(C) differential responses temperature；(D) the differential responses time.
Fig. 6 (A) is the scanning electron microscope (SEM) photograph of different temperatures synthesis Au@AuPtNPs of the present invention；(B) it is different temperatures of the present invention
The size statistical analysis of synthesis Au@AuPtNPs.
Fig. 7 is stability change figure of the Au AuPtNPs catalase activities obtained in the embodiment of the present invention 1 with the time.
Fig. 8 is the enzymatic activity comparing result figure of Au@AuPtNPs obtained in the embodiment of the present invention 1 and catalase, wherein
Michaelis constant (K during including with hydrogen peroxide as substratem), maximum reaction rate (Vmax) and catalytic constant (Kcat)。
Specific embodiment
Technical scheme is further detailed and described below by way of specific embodiment combination accompanying drawing.
Embodiment 1
The technology path of the present embodiment as shown in figure 1, specific experiment condition gropes process as shown in Figure 5 and Figure 6, specific step
It is rapid as follows：
(1) 100mL 0.01wt% gold chlorides are added in 250mL round-bottomed flasks, lower boiling reflux is continuously stirred, at the uniform velocity
The reduction of sodium citrate gold chloride of 1mL 3wt% is added to form 13nm gold nano grains (such as Fig. 3).In experimentation, solution by
It is faint yellow to be changed into black, then become red.
(2) under conditions of 400rpm stirrings, 1.6mM chlorine platinum is contained to 10mL is added in 100mL 13nm AuNPs solution
Acid and the mixed liquor of 8mM gold chlorides.Then, 5mL 80mM ascorbic acid, room temperature 400rpm stirring 8h synthesis Au@are slowly added to
AuPtNPs, i.e., the nano particle of described strong catalase activity.With ultraviolet-visible spectrophotometer (UV-vis), scanning electricity
Mirror (SEM), transmission electron microscope (TEM), high-resolution-ration transmission electric-lens (HR-TEM) and x-ray photoelectron power spectrum (XPS) analyze synthetic
Au@AuPtNPs, as a result as shown in Fig. 2,3,4 and 6.
(3) Au@AuPtNPs catalase activities are determined：100 μ L 0.1M H are added in 200 μ L centrifuge tubes2O2With 2 μ
L 2.5nM Au@AuPtNPs, upside down reacts 10min, nano particle catalysis H at 37 DEG C after mixing2O2Decompose what is produced
Oxygen concentrates on centrifuge tube top.Pipettor sample volume is adjusted to 100 μ L, centrifuge tube upper end is siphoned away completely with pipettor
The water that density is 1g/mL is drawn after oxygen again, the volume of the water outlet that can just be converted using the quality of balance measure pipettor extract water.
Au@AuPtNPs nano particles catalysis H is calculated using drainage2O2The volume for producing oxygen is decomposed, as a result as shown in FIG. 7 and 8.
Those of ordinary skill in the art understand, when composition of the invention and parameter change in following ranges, still are able to
To technique effect same as the previously described embodiments or close, protection scope of the present invention is come under：
A kind of preparation method of the nano particle of strong catalase activity, comprises the following steps：
(1) by chlorauric acid solution boiling reflux with continuous stirring, while at the uniform velocity adding sodium citrate solution to reduce chlorine
Auric acid forms particle diameter 12~14nm gold nano grains, and wherein chlorauric acid solution and the volume ratio of sodium citrate solution is 100:1~
Gold chloride containing 0.01~0.02wt% in 1.5, and chlorauric acid solution, contains 2.8~3.5wt%'s in sodium citrate solution
Sodium citrate；
(2) to the mixed solution that gold chloride and chloroplatinic acid are added in material obtained in step (1), it is then slowly added into anti-bad
Hematic acid solution, with the speed 2~12h of stirring reaction of 100~800rpm at a temperature of 10~50 DEG C, so that above-mentioned gold nano grain
The upper golden platinum bimetallic shell of parcel, obtains final product the nano particle of the strong catalase activity, wherein in the reactant of the step
In system, the concentration of chloroplatinic acid is 0.04~0.064mM, and the concentration of gold chloride is 0.2~3.2mM, the concentration of ascorbic acid for 1~
16mM, and the mass ratio of above-mentioned chloroplatinic acid and gold chloride is 10~40:100, the gross mass and Vitamin C of chloroplatinic acid and gold chloride
The mass ratio of acid is 13.4~53.5:100, the mass ratio of gold nano grain and chloroplatinic acid and gold chloride gross mass for 2.7~
11.2:100。
Preferably, in the step (2), temperature is 20~40 DEG C, and mixing speed is 300~500rpm, and the reaction time is 5
~9h, in reaction system：The concentration of chloroplatinic acid is 0.08~0.32mM, and the concentration of gold chloride is 0.4~1.6mM, ascorbic acid
Concentration be 2~8mM.
The above, only presently preferred embodiments of the present invention, therefore can not according to this limit the scope of present invention implementation, i.e.,
The equivalence changes made according to the scope of the claims of the present invention and description and modification, all should still belong in the range of the present invention covers.
Claims (7)
1. a kind of preparation method of the nano particle of strong catalase activity, it is characterised in that：Comprise the following steps：
(1) by chlorauric acid solution boiling reflux with continuous stirring, while at the uniform velocity adding sodium citrate solution to reduce gold chloride
Particle diameter 12~14nm gold nano grains are formed, wherein chlorauric acid solution and the volume ratio of sodium citrate solution is 100:1~1.5,
And the gold chloride containing 0.01~0.02wt% in chlorauric acid solution, the lemon containing 2.8~3.5wt% in sodium citrate solution
Sour sodium；
(2) to the mixed solution that gold chloride and chloroplatinic acid are added in material obtained in step (1), it is then slowly added into ascorbic acid
Solution, with the speed 2~12h of stirring reaction of 100~800rpm at a temperature of 10~50 DEG C, so as to be wrapped on above-mentioned gold nano grain
Golden platinum bimetallic shell is wrapped up in, the nano particle of the strong catalase activity is obtained final product, wherein in the reaction system of the step,
The concentration of chloroplatinic acid is 0.04~0.064mM, and the concentration of gold chloride is 0.2~3.2mM, and the concentration of ascorbic acid is 1~16mM,
And the mass ratio of above-mentioned chloroplatinic acid and gold chloride is 10~40:100, the matter of the gross mass and ascorbic acid of chloroplatinic acid and gold chloride
Amount is than being 13.4~53.5:100, gold nano grain is 2.7~11.2 with the mass ratio of chloroplatinic acid and gold chloride gross mass:100.
2. a kind of preparation method of the nano particle of strong catalase activity as claimed in claim 1, it is characterised in that：Institute
Stating step (1) is：By chlorauric acid solution boiling reflux with continuous stirring, while at the uniform velocity adding sodium citrate solution to reduce chlorine
Auric acid forms particle diameter 13nm gold nano grains, and wherein chlorauric acid solution and the volume ratio of sodium citrate solution is 100:1, and chlorine gold
Gold chloride containing 0.01wt% in acid solution, the sodium citrate containing 3wt% in sodium citrate solution.
3. a kind of preparation method of the nano particle of strong catalase activity as claimed in claim 1 or 2, its feature exists
In：Temperature in the step (2) is 20~40 DEG C.
4. a kind of preparation method of the nano particle of strong catalase activity as claimed in claim 1 or 2, its feature exists
In：Mixing speed in the step (2) is 300~500rpm.
5. a kind of preparation method of the nano particle of strong catalase activity as claimed in claim 1 or 2, its feature exists
In：Reaction time in the step (2) is 5~9h.
6. a kind of preparation method of the nano particle of strong catalase activity as claimed in claim 1 or 2, its feature exists
In：The concentration of gold chloride is 0.4~1.6mM in the step (2).
7. a kind of preparation method of the nano particle of strong catalase activity as claimed in claim 1 or 2, its feature exists
In：The concentration of ascorbic acid is 2~8mM in the step (2).
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| CN109382523A (en) * | 2018-11-05 | 2019-02-26 | 华中科技大学 | A kind of preparation method of the alloy hollow nano material with catalase activity |
| CN110960696B (en) * | 2019-12-02 | 2021-12-24 | 西南大学 | Preparation method of hollow cobalt oxide @ gold platinum nanospheres with biological enzyme activity |
| CN111702186B (en) * | 2020-07-23 | 2023-03-24 | 苏州大学 | Preparation method of gold nanoparticles with adjustable size |
| CN113664198B (en) * | 2021-08-24 | 2023-02-24 | 杭州微净科技有限公司 | Rapid chemical coating method for compact gold nanoparticles and production equipment thereof |
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