CN108912375A - The oriented growth nanogold bacteria cellulose/compound film method of carbon pipe - Google Patents
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- CN108912375A CN108912375A CN201810550612.6A CN201810550612A CN108912375A CN 108912375 A CN108912375 A CN 108912375A CN 201810550612 A CN201810550612 A CN 201810550612A CN 108912375 A CN108912375 A CN 108912375A
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Abstract
The invention discloses oriented growth nanogold bacteria cellulose/compound film methods of carbon pipe, the described method comprises the following steps:Culture acetobacter xylinum obtains BC film, film process is clean and be ground into cotton-shaped;MWCNTs solution is mixed with cotton-shaped BC film, obtains BC/MWCNTs composite membrane through filtering;BC/MWCNTs/Au NPs composite base material is obtained with PEI solution and chlorauric acid solution processing gained composite membrane to fix nano Au particle on its surface;BC/MWCNTs/Au NPs composite base material is immersed in the absorption that enzyme is carried out in laccase and glucolase solution.Using BC film as electrode material matrix, the adsorption program of MWCNTs is simplified, while BC has good biocompatibility and it is made to have extensive biologic applications prospect;Common filter is mixed with MWCNTs solution using crushing BC film and obtains the technical treatment mode of composite membrane, so that the carbon pipe of composite film surface is evenly distributed, electric conductivity is good;In BC/MWCNTs composite film surface controllable growth nanogold, the response sensitivity of sensor is improved using the electron transmission effect of nanogold and concerted catalysis effect.
Description
Technical field
The invention belongs to biological fuel cell technical fields, are related to biological fuel cell and the preparation of self energizing biosensor
Method, specially oriented growth nanogold bacteria cellulose/compound film method of carbon pipe.
Background technique
Biological fuel cell is generally divided into microbiocidal and two kinds of enzyme type, and wherein enzymatic biological fuel battery application prospect is more
Add wide.Fuel (common are glucose oxidase, cellobiose dehydrogenase, Fructose dehydrogenase, ethyl alcohol in anode enzyme catalyst
Dehydrogenase etc.) under the action of be oxidized, the electronics of generation reaches cathode by external circuit, and proton is reached negative by proton exchange membrane
Pole, oxide (generally oxygen) obtain electronics in cathode and are reduced (product is generally water), and whole process forms an access,
Constitute enzymatic biological fuel battery.
The design principle of self energizing biosensor is derived mainly from biological fuel cell.As long as studies have shown that solving life
The problem of object compatibility, the self energizing biosensor based on glucose fuel continuously monitor people as implanted blood glucose sensor
Internal blood glucose amount variation is practicable.
A kind of bacteria cellulose superfine nano fiber similar with plant cellulose as chemical property possesses many excellent
Physical and chemical performance, such as big specific surface area, high crystallinity, good mechanical performance and specific retention.And by bacterium fibre
Dimension element is formed by natural cellulose membrane with good flexibility and good biocompatibility, has been widely used in giving birth to
Object medical treatment aspect, such as artificial skin, artificial blood vessel and wound dressing etc..By bacteria cellulose, (such as carbon is received with conductive materials
Mitron, graphene, conduction high polymer) combination can assign its excellent electric conductivity, thus as flexible electrode apply lithium from
Sub- battery, supercapacitor and sensor etc..
On BC/CNT membrane electrode surface, controllable growth nanogold particle can be improved the sensitivity of sensor and regulate and control its inspection
Linear range.Nanogold possesses good biocompatibility, and enzyme active center and interelectrode electronics can be improved in nanogold
Transfer efficiency and then the response sensitivity for enhancing biosensor.Some researches show that can by change environment temperature, pH,
The means such as halide ion are added to regulate and control the appearance and size in the nanogold of bacteria cellulose surface growth.
Summary of the invention
The technical issues of solution:In order to overcome the drawbacks of the prior art, current self energizing biological sensor electrode spirit is solved
The problem that sensitivity is poor, biocompatibility is low, the present invention provides oriented growth nanogold bacteria cellulose/carbon pipe composite membrane sides
Method.
Technical solution:The oriented growth nanogold bacteria cellulose/compound film method of carbon pipe, the method includes following steps
Suddenly:
Step 1, culture acetobacter xylinum obtain BC film, film process is clean and be ground into cotton-shaped;
Step 2 mixes MWCNTs solution with cotton-shaped BC film, obtains BC/MWCNTs composite membrane through filtering;
Step 3 is obtained with PEI solution and chlorauric acid solution processing gained composite membrane with fixing nano Au particle on its surface
To BC/MWCNTs/Au NPs composite base material;
BC/MWCNTs/Au NPs composite base material is immersed in the suction that enzyme is carried out in laccase and glucolase solution by step 4
It is attached.
The abbreviation or abbreviation being related to are explained as follows:BC is Bacterial Cellulose, i.e. bacteria cellulose film;
MWCNTs is Multi-walled Carbon nanotubes, i.e. multiple-wall carbon nanotube;PEI is polyetherimide.
Preferably, cultivating the step of acetobacter xylinum obtains BC film is:Yeast is soaked into bacterium, tryptone, mannitol and is dissolved in water,
Acetobacter xylinum fermentation culture, autoclave sterilization are prepared, aseptic inoculation is placed in biochemical cultivation case and cultivates;By upgrowth situation
Good BC film is handled, and to wash away a large amount of impurity and bacterial nutrientss of film surface, processing, which is placed in distilled water, impregnates
It is spare.
Preferably, the step of BC/MWCNTs compound film preparation is:BC film is crushed into cotton-shaped, preparation MWCNTs solution, will
MWCNTs solution is added in the BC film mixed liquor of cotton-shaped dispersion, stirs evenly, and the BC/ of carbon pipe uniform adsorption is made through filtering
MWCNTs composite membrane.
Preferably, the step of BC/MWCNTs/Au NPs compound film preparation is:Chlorauric acid solution, PEI solution are prepared, will be walked
Suddenly the BC/MWCNTs composite membrane that (2) obtain is immersed in PEI solution, is added chlorauric acid solution after stewing process, in baking oven
Reason, natural air drying obtains BC/MWCNTs/Au NPs composite base material after taking-up.
Preferably, the processing step of BC/MWCNTs/Au NPs composite base material is:Prepare laccase and glucose oxidase
BC/MWCNTs/Au NPs composite base material is immersed in laccase and glucose oxidase aqueous solution, makes laccase and grape by aqueous solution
Carbohydrate oxidase is individually fixed on BC/MWCNTs/Au NPs substrate, and BC/MWCNTs/Au NPs composite membrane is made.
Preferably, acetobacter xylinum fermented and cultured formula of liquid is:Yeast soaks bacterium 3g/L, tryptone 5g/L, mannitol 25g/
L;Autoclave sterilization temperature is 120 DEG C;Condition of culture is after connecing bacterium:It is cultivated in 30 DEG C of biochemical cultivation case.
Preferably, the specific method of BC film process is:After carrying out repeated flushing with deionized water, the NaOH for being placed in 4g/L is molten
Liquid handles 2h in 80 DEG C of water-baths, to wash away a large amount of impurity and bacterial nutrientss of film surface.
Preferably, MWCNTs solution concentration is 1mg/mL, prepares and micro surfactant is added in the solution processes, make
Solid dispersion is more uniform.
Preferably, chlorauric acid solution and PEI solution concentration are respectively 20mmol/L and 10g/L, and specific processing method is:It takes
PEI solution submerges BC/MWCNTs film, stands 30min, the PEI solution that the chlorauric acid solution volume of addition is 0.5 times at room temperature
Volume, is placed in 60 DEG C of baking oven dryings 1 hour.
Preferably, laccase and glucose oxidase solution concentration are 1mg/mL, inhale laccase and glucose oxidase sufficiently
The processing method of attached fixation is:After composite membrane is immersed laccase and glucose oxidase solution, it is placed in 8h in 4 DEG C of refrigerators.
Oriented growth nanogold bacteria cellulose of the present invention/carbon pipe compound film method principle is:Using BC as
Basis material introduces MWCNTs, is deposited on MWCNTs effectively in its nanoporous network structure, is uniformly distributed with realizing.With
Au NPs is prepared in situ as reducing agent in PEI, while BC can be assisted to play the role of stable Au NPs, prevents it from assembling.
Beneficial effect:(1) for the present invention using BC film as electrode material matrix, active group simplifies MWCNTs's
Adsorption program, while BC has good biocompatibility and it is made to have extensive biologic applications prospect;(2) present invention adopts
Common filter, which is mixed, with MWCNTs solution with crushing BC film obtains the technical treatment mode of composite membrane, so that composite film surface
Carbon pipe is evenly distributed, and electric conductivity is good;(3) present invention is in BC/MWCNTs composite film surface controllable growth nanogold, using receiving
The electron transmission effect of meter Jin and concerted catalysis effect improve the response sensitivity of sensor.
Detailed description of the invention
Fig. 1 is the scanning electron microscope (SEM) photograph of BC/MWCNTs/Au NPs film, and Fig. 1 (a) is 20 times of amplification, and Fig. 1 (b) is amplification 50
Times;
Fig. 2 is the infrared spectrogram of BC film, BC/MWCNTs film and BC/MWCNTs/Au NPs film;
Fig. 3 is the thermogravimetric analysis figure of BC film, BC/MWCNTs film and BC/MWCNTs/Au NPs film;
Fig. 4 is BC film, BC/MWCNTs film and BC/MWCNTs/Au NPs film XRD spectrum;
Fig. 5 is the electrochemical properties test result figure of laccase electrode, and Fig. 5 A is that BC/MWCNTs/Au NPs-Lac electrode exists
Sweep speed is respectively 50,100,150,200,250,300 in (pH 4.5) in 0.1M acetic acid/sodium acetate buffer,
The cyclic voltammetry curve figure of 350mV/s, Fig. 5 B are anode and peak cathode current to the calibration curve for sweeping speed;Lac is laccase;
Fig. 6 is the electrochemical properties test result figure of glucose oxidase electrode, and Fig. 6 A is BC/MWCNTs/Au NPs-
Sweep speed is respectively 50 to GOx electrode in (pH 4.5) in 0.1M acetic acid/sodium acetate buffer containing 50mM glucose,
100,150,200,250,300mV/s cyclic voltammetry curve figure, Fig. 6 B are the calibration of anode and peak cathode current to speed is swept
Curve;GOx is glucose oxidase.
Specific embodiment
Following embodiment further illustrates the contents of the present invention, but should not be construed as limiting the invention.Without departing substantially from
In the case where spirit of that invention and essence, to modification made by the method for the present invention, step or condition and replaces, belong to the present invention
Range.Unless otherwise specified, the conventional means that technological means used in embodiment is well known to those skilled in the art.
Embodiment 1
The oriented growth nanogold bacteria cellulose/compound film method of carbon pipe, the described method comprises the following steps:
Step 1, culture acetobacter xylinum obtain BC film, film process is clean and be ground into cotton-shaped;
Step 2 mixes MWCNTs solution with cotton-shaped BC film, obtains BC/MWCNTs composite membrane through filtering;
Step 3 is obtained with PEI solution and chlorauric acid solution processing gained composite membrane with fixing nano Au particle on its surface
To BC/MWCNTs/Au NPs composite base material;
BC/MWCNTs/Au NPs composite base material is immersed in the suction that enzyme is carried out in laccase and glucolase solution by step 4
It is attached.
The abbreviation or abbreviation being related to are explained as follows:BC is Bacterial Cellulose, i.e. bacteria cellulose film;
MWCNTs is Multi-walled Carbon nanotubes, i.e. multiple-wall carbon nanotube;PEI is polyetherimide.
Cultivating the step of acetobacter xylinum obtains BC film is:Yeast is soaked into bacterium, tryptone, mannitol and is dissolved in water, preparation wood
Acetobacter fermentation culture, autoclave sterilization, aseptic inoculation are placed in biochemical cultivation case and cultivate;Upgrowth situation is good
BC film is handled, to wash away a large amount of impurity and bacterial nutrientss of film surface, processing be placed in distilled water impregnate it is spare.
The step of BC/MWCNTs compound film preparation is:BC film is crushed into cotton-shaped, preparation MWCNTs solution, by MWCNTs
Solution is added in the BC film mixed liquor of cotton-shaped dispersion, stirs evenly, and the BC/MWCNTs through filtering obtained carbon pipe uniform adsorption is compound
Film.
The step of BC/MWCNTs/Au NPs compound film preparation is:Chlorauric acid solution, PEI solution are prepared, step (2) are obtained
To BC/MWCNTs composite membrane be immersed in PEI solution, chlorauric acid solution is added after stewing process, is handled in baking oven, after taking-up
Natural air drying obtains BC/MWCNTs/Au NPs composite base material.
The processing step of BC/MWCNTs/Au NPs composite base material is:The aqueous solution of laccase and glucose oxidase is prepared,
BC/MWCNTs/Au NPs composite base material is immersed in laccase and glucose oxidase aqueous solution, laccase and glucose oxidase are made
It is individually fixed on BC/MWCNTs/Au NPs substrate, BC/MWCNTs/Au NPs composite membrane is made.
Acetobacter xylinum fermented and cultured formula of liquid is:Yeast soaks bacterium 3g/L, tryptone 5g/L, mannitol 25g/L;High temperature is high
Pressing sterilising temp is 120 DEG C;Condition of culture is after connecing bacterium:It is cultivated in 30 DEG C of biochemical cultivation case.
The specific method of BC film process is:After carrying out repeated flushing with deionized water, the NaOH solution of 4g/L is placed in 80
2h is handled in DEG C water-bath, to wash away a large amount of impurity and bacterial nutrientss of film surface.
MWCNTs solution concentration is 1mg/mL, prepares and micro surfactant is added in the solution processes, makes solid point
It dissipates more uniform.
Chlorauric acid solution and PEI solution concentration are respectively 20mmol/L and 10g/L, and specific processing method is:Take PEI solution
BC/MWCNTs film is submerged, stands 30min at room temperature, the PEI liquor capacity that the chlorauric acid solution volume of addition is 0.5 times, and
It is placed in 60 DEG C of baking oven dryings 1 hour.
Laccase and glucose oxidase solution concentration are 1mg/mL, and laccase and glucose oxidase is made sufficiently to adsorb fixation
Processing method is:After composite membrane is immersed laccase and glucose oxidase solution, it is placed in 8h in 4 DEG C of refrigerators.
The oriented growth nanogold bacteria cellulose/carbon pipe composite membrane prepared is detected, it is as a result as follows:
As shown in Figure 1, Fig. 1 (a) shows that carbon nanotube and nano Au particle distribution are relatively uniform, carbon nanotube is tightly wrapped
It is rolled in the bacteria cellulose with ultra micro gap structure, while passing through distribution of the gold nanoparticle on BC/MWCNTs carrier,
Also explanation achieves preferable effect using the fixed carbon nanotube of suction method and in the fixed nanogold of composite film surface.By (b)
Figure is as it can be seen that bacteria cellulose shows the tridimensional network of random orientation, and surface pore is not of uniform size, elongated carbon nanotube
Be filled between the fiber gap of bacteria cellulose at random, a large amount of Au by uniform adsorption in composite film surface, to improve electrochemistry spirit
Quick property provides necessary condition.
As shown in Fig. 2, in the stretching vibration region of C-O, 1659cm-1And 1061cm-1Absorption peak to respectively correspond C-O non-
Symmetrical stretching vibration and pyranose ring skeletal vibration, 2973cm-1Place is the 3465cm as caused by the vibration of-CH--1Place be by-
Caused by OH group symmetrical stretching vibration.As seen from the figure, after multi-walled carbon nanotube being added, spectrogram peak is in 2973cm-1Locate micro-
Amount enhancing, in 3465cm-1Place is remarkably reinforced, this is because the addition of multi-walled carbon nanotube is so that flexible caused by the superposition of-CH- key
Vibration enhancing, while a large amount of-OH groups are introduced, illustrate there was only Physical interaction between BC and multi-walled carbon nanotube;Load is received
Meter Jin Hou, each wave crest is without significant change, then supported nano-gold particle does not generate BC/MWCNTs film items chemical property substantially
It influences.
As shown in figure 3, the ingredient of BC is cellulose, its thermal decomposition occurs in 220~400 DEG C of temperature ranges, this single order
The mass loss rate of section sample is up to 75%;It can see BC there are two apparent weightless peak from DTG curve, respectively correspond 220~
270 DEG C of temperature sections and 300~400 DEG C of temperature sections, caused by the two weightless peaks are all BC thermal decomposition.Wherein 300~400 DEG C it is right
The weightless peak-to-peak value answered is larger, illustrates that BC mass loss is very fast in this stage, and when temperature is 320 DEG C or so, BC heat point
Solution rate reaches maximum.The thermal decomposition of BC/MWCNTs film occurs in 350~400 DEG C of temperature ranges, the quality of this stage sample
Loss late about 50%;Can see BC/MWCNTs from DTG curve has an apparent weightless peak, corresponding 350~400 DEG C of temperature
Section.The thermal decomposition of BC/MWCNTs/Au NPs film occurred in 190~250 DEG C and 350~400 DEG C of temperature ranges, the first and second stage
The mass loss rate of sample respectively may be about 5% and 20%;DTG curve shows that first stage weightless peak is smaller, and mass loss is slower,
First stage is the main mass loss stage.
Comparison can obtain, and compare pure BC film, and the beginning weightless temperature of BC/MWCNTs film improves about 100 DEG C, and percent weight loss reduces
About 25%;BC/MWCNTs/Au NPs film beginning weightless temperature then reduces about 30 DEG C, but main weightless temperature improves about 50 DEG C, and loses
Weight percentage substantially reduces.It to sum up states, BC film thermal stability after processing improves.
As shown in figure 4, comparing pure BC film, the diffraction of the bacteria cellulose of BC/MWCNTs and BC/MWCNTs/Au NPs film
Peak heights increase, and diffracted intensity significantly improves, then the crystallization degree of bacteria cellulose film after processing increases;Meanwhile in BC/
On MWCNTs and BC/MWCNTs/Au NPs film, there is peak at (004) in C (002), C (100), C (101), C, it was demonstrated that carbon is received
Mitron bacteria cellulose film adsorption in order.Compared to BC/CNT film, the film after loading nanometer Au particle is in Au
(111), there is peak at (220) in Au (200), Au, illustrate absorption of nanometer Au on film in order.
As shown in figure 5, it can be observed that apparent anode and cathode peak point current, and with the increase of sweep speed, electricity
The peak value of stream is positive respectively, inversely increases, and the AuNPs and BC/MWCNTs that this explanation is connect with electrode surface on BC can be into
The transfer of row laccase redox reaction electronics.As shown in Figure 5 B, using anode peak and cathodic peak current as in 50-350mV model
Enclose the subduplicate function of interior sweep speed, it is seen that with the raising for sweeping speed, linear increase is presented in peak point current.The result shows that
The electrochemical reaction on laccase electrode surface has controllability, for stable electrochemical system.
As shown in fig. 6, in fig. 6, it has also been observed that apparent anode and cathode peak point current, sweep speed by
50mV increases to 300mV, and anode peak current is gradually increased to about 0.7mA by 0.2mA, cathodic peak current value by -0.2mA gradually
About -0.83mA is increased to, illustrates to couple between glucose oxidase and BC/MWCNTs/AuNPs bioelectrode good.Such as Fig. 6 B
It is shown, it is same to advise using anode peak and cathodic peak current as the subduplicate function of the sweep speed within the scope of 50-350mV
Rule, with the raising for sweeping speed, linear increase is presented in peak point current.The result shows that glucose oxidase is modified BC/MWCNTs/
The chemical reaction of AuNPs electrode surface has controllability.
Claims (10)
1. the oriented growth nanogold bacteria cellulose/compound film method of carbon pipe, which is characterized in that the method includes following steps
Suddenly:
Step 1, culture acetobacter xylinum obtain BC film, film process is clean and be ground into cotton-shaped;
Step 2 mixes MWCNTs solution with cotton-shaped BC film, obtains BC/MWCNTs composite membrane through filtering;
Step 3 is obtained with PEI solution and chlorauric acid solution processing gained composite membrane with fixing nano Au particle on its surface
BC/MWCNTs/Au NPs composite base material;
BC/MWCNTs/Au NPs composite base material is immersed in the absorption that enzyme is carried out in laccase and glucolase solution by step 4.
2. the oriented growth nanogold bacteria cellulose according to claim 1/compound film method of carbon pipe, which is characterized in that
Cultivating the step of acetobacter xylinum obtains BC film is:Yeast is soaked into bacterium, tryptone, mannitol and is dissolved in water, prepares acetobacter xylinum hair
Ferment culture solution, autoclave sterilization, aseptic inoculation are placed in biochemical cultivation case and cultivate;The good BC film of upgrowth situation is carried out
Processing, to wash away a large amount of impurity and bacterial nutrientss of film surface, processing be placed in distilled water impregnate it is spare.
3. the oriented growth nanogold bacteria cellulose according to claim 1/compound film method of carbon pipe, which is characterized in that
The step of BC/MWCNTs compound film preparation is:BC film be crushed into it is cotton-shaped, prepare MWCNTs solution, MWCNTs solution is added
It in the BC film mixed liquor of cotton-shaped dispersion, stirs evenly, the BC/MWCNTs composite membrane of carbon pipe uniform adsorption is made through filtering.
4. the oriented growth nanogold bacteria cellulose according to claim 1/compound film method of carbon pipe, which is characterized in that
The step of BC/MWCNTs/Au NPs compound film preparation is:Prepare chlorauric acid solution, PEI solution, the BC/ that step (2) is obtained
MWCNTs composite membrane is immersed in PEI solution, and chlorauric acid solution is added after stewing process, handles in baking oven, natural wind after taking-up
It is dry to obtain BC/MWCNTs/Au NPs composite base material.
5. the oriented growth nanogold bacteria cellulose according to claim 1/compound film method of carbon pipe, which is characterized in that
The processing step of BC/MWCNTs/Au NPs composite base material is:The aqueous solution for preparing laccase and glucose oxidase, by BC/
MWCNTs/Au NPs composite base material immerses in laccase and glucose oxidase aqueous solution, distinguishes laccase and glucose oxidase
It is fixed on BC/MWCNTs/Au NPs substrate, BC/MWCNTs/Au NPs composite membrane is made.
6. the oriented growth nanogold bacteria cellulose according to claim 2/compound film method of carbon pipe, which is characterized in that
Acetobacter xylinum fermented and cultured formula of liquid is:Yeast soaks bacterium 3g/L, tryptone 5g/L, mannitol 25g/L;Autoclave sterilization temperature
Degree is 120 DEG C;Condition of culture is after connecing bacterium:It is cultivated in 30 DEG C of biochemical cultivation case.
7. the oriented growth nanogold bacteria cellulose according to claim 2/compound film method of carbon pipe, which is characterized in that
The specific method of BC film process is:After carrying out repeated flushing with deionized water, the NaOH solution of 4g/L is placed in 80 DEG C of water-baths
Middle processing 2h, to wash away a large amount of impurity and bacterial nutrientss of film surface.
8. the oriented growth nanogold bacteria cellulose according to claim 3/compound film method of carbon pipe, which is characterized in that
MWCNTs solution concentration is 1mg/mL.
9. the oriented growth nanogold bacteria cellulose according to claim 4/compound film method of carbon pipe, which is characterized in that
Chlorauric acid solution and PEI solution concentration are respectively 20mmol/L and 10g/L, and specific processing method is:Take PEI solution by BC/
The submergence of MWCNTs film, stands 30min at room temperature, and the PEI liquor capacity that the chlorauric acid solution volume of addition is 0.5 times is placed in
60 DEG C baking oven drying 1 hour.
10. the oriented growth nanogold bacteria cellulose according to claim 5/compound film method of carbon pipe, which is characterized in that
Laccase and glucose oxidase solution concentration are 1mg/mL, and laccase and glucose oxidase is made sufficiently to adsorb fixed processing method
It is:After composite membrane is immersed laccase and glucose oxidase solution, it is placed in 8h in 4 DEG C of refrigerators.
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CN112094437A (en) * | 2019-06-18 | 2020-12-18 | 南京理工大学 | Preparation method of bacterial cellulose/polyaniline/gold nanoparticle composite material |
CN111517475A (en) * | 2020-04-30 | 2020-08-11 | 宣宇青 | Method for degrading chlorophenol pollutants in water body by utilizing POPs (polymer-organic compounds) reduction material |
CN113372620A (en) * | 2021-05-27 | 2021-09-10 | 东华大学 | Photo-thermal conversion material with nano heterostructure and preparation method and application thereof |
CN113372620B (en) * | 2021-05-27 | 2022-05-31 | 东华大学 | Photo-thermal conversion material with nano heterostructure and preparation method and application thereof |
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