CN108760858A - A kind of nano titania column array enzyme electrode and its preparation method and application of nanometer of modified by silver - Google Patents
A kind of nano titania column array enzyme electrode and its preparation method and application of nanometer of modified by silver Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
- G01N27/3272—Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/308—Electrodes, e.g. test electrodes; Half-cells at least partially made of carbon
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
- G01N27/3277—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction being a redox reaction, e.g. detection by cyclic voltammetry
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
- G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles
Abstract
The present invention relates to the nano titania column array enzyme electrodes and its preparation method and application of a kind of nanometer of modified by silver, and the method grows TiO on carbon paper first using carbon paper as base material2Nano column array；Then growth there is into TiO2The carbon paper of nano column array is immersed in silver sol 3-15 hours, takes out, the nano titania column array of nanometer modified by silver is drying to obtain after being cleaned with deionized water；Finally with bovine serum albumin-glutaraldehyde as cross linker, glucose oxidase is fixed on to the nano titania column array surface of nanometer modified by silver using improvement cross-linking method, that is, forms the nano titania column array enzyme electrode of nanometer modified by silver.The nano titania column array enzyme electrode of nanometer modified by silver prepared by the present invention has better electro catalytic activity, can be advantageously applied in glucose biological sensor or enzyme thermistor devices.
Description
Technical field
The present invention relates to biosensor technology field more particularly to the nano titania column battle arrays of a kind of nanometer of modified by silver
Row enzyme electrode and its preparation method and application.
Background technology
Glucose oxidase electrode is a kind of to measure the biosensor of glucose in solutions content to be measured, in base
Fixed enzyme membrane is housed, when electrode is inserted into solution to be measured, the glucose oxidase in enzyme membrane occurs on the sensitive area of plinth electrode
Catalysis reaction generates electrode active material, and response current changes with the concentration of active material, and it is glycoxidative thus to measure the grape
The concentration of reactant or reaction product in the reaction that enzyme is catalyzed, this glucose oxidase electrode biosensor have structure
Simply, high sensitivity, selectivity are good and can real-time monitoring and other advantages, it has also become the hot spot of research.
Composition, surface texture and the glucose oxidase of electrode material are to determine grape glycosyloxy in the fixation of electrode surface
An important factor for changing enzyme electrode biosensor electro catalytic activity.Nano structural material has big specific surface area, can be Portugal
The fixed of grape carbohydrate oxidase provides more active sites, to improve the fixed efficiency of glucose oxidase, enhances glucose
The electro catalytic activity of oxidase electrode biosensor.Current various nano materials have been used for glucose oxidase electricity
In extremely, such as carbon nano-particle, Au nano particles, Pt nano particles and SiO2、Al2O3、MnO2And TiO2Equal oxidate nanos
Grain.Wherein, due to TiO2Nano material has good biocompatibility, relatively high electric conductivity, nontoxic, stable and valence
The advantages that lattice are cheap, the TiO of various structures2Nano material has been used in glucose enzyme electrode biosensor preparation process
The fixation of enzyme.
Compared to other structures, the TiO of high-sequential2Nano-array can provide half-duplex channel for electron-transport, and electronics passes
Lead efficient, TiO at present2Nano-tube array has been widely used as the carrier material of glucose oxidase.However, due to surface
The effect of power, either glucose oxidase electrode surface diffusion to electrode surface of fixation or electrolyte solution, all
It is difficult to reach TiO2The inside of nanotube, thus TiO cannot be given full play to2It the advantages of nano-tube array large specific surface area, affects
The raising of the fixed efficiency of glucose oxidase.In addition, merely by TiO2Nano material constitute enzyme electrode also often show compared with
Low bioelectrocatalytic activity.
Invention content
For the above problem of the prior art, the object of the present invention is to provide a kind of nanometers with more preferable electrocatalysis characteristic
The nano titania column array enzyme electrode of modified by silver and the preparation method and application of the enzyme electrode.
In order to solve the above-mentioned technical problem, the first aspect of the present invention provides the nano titania of a kind of nanometer of modified by silver
The preparation method of column array enzyme electrode, this approach includes the following steps：
(1) using carbon paper as base material, TiO is grown on the carbon paper2Nano column array；
(2) growth in step (1) there is into TiO2The carbon paper of nano column array is immersed in silver sol 3-15 hours, is then taken
Go out, the nano titania column array of nanometer modified by silver is drying to obtain after being cleaned with deionized water；
(3) with bovine serum albumin-glutaraldehyde as cross linker, glucose oxidase is fixed on step using cross-linking method is improved
(2) the nano titania column array surface of the nanometer modified by silver described in, that is, form the titanium dioxide of the nanometer modified by silver
Nano column array enzyme electrode.
The present invention uses TiO2Nano column array as enzyme electrode carrier material, due to TiO2Nano-tube array is compared,
TiO2Nano column array not only equally has big specific surface area and high electronics conduction efficiency, but also it also helps electrolyte
Diffusion of the solution to electrode surface, and by TiO2The surface finish nano Argent grain of nano column array, to greatly enhance
Its electro catalytic activity.
Using carbon paper as base material, carbon paper is bonded the present invention by carbon fiber skeleton and carbon material, conductive
Well, the advantages that property is stable and cheap.Carbon paper surface is coarse, internal there are a large amount of holes, therefore the ratio with bigger
Surface is growth TiO2The ideal base material of nano column array.
Further, described using carbon paper as base material in step (1), TiO is grown on carbon paper2Nano column array
Specifically include following steps：
(1.1) carbon paper of certain size size is taken, it is spare after cleaning, drying；
(1.2) it is 1 according to volume ratio by titanium source and absolute ethyl alcohol:4~1:20 mixing, then drip into the mixed solution
The volume ratio of ice acetic acid, the glacial acetic acid and titanium source is 1:50~1:250, it is stirred to react to obtain TiO at room temperature2Colloid is molten
Liquid；
(1.3) TiO being immersed in the carbon paper being dried for standby in step (1.1) in step (1.2)2In colloidal solution, so
Drying is taken out afterwards, and 10~30min postcoolings are handled at a temperature of 300~400 DEG C to room temperature, obtain absorption TiO2Nano particle
Carbon paper；
(1.4) by the absorption TiO2The carbon paper of nano particle is put into hydrochloric acid and the mixed liquor of titanium source, 150~200
It is reacted 4~20 hours at DEG C, product obtained by the reaction is washed off dry after the reaction solution of remained on surface with deionized water, is obtained
Growth has unformed TiO2The carbon paper of nano column array；
(1.5) product after will be dry in step (1.4), which is placed in the high temperature furnace of inert gas atmosphere, carries out calcination processing,
Calcination temperature is 400~600 DEG C, and calcination time is 1~3 hour, and heating rate is 3~10 DEG C/min, and then control cooling is
Must grow has the TiO of crystal phase2The carbon paper of nano column array.
Further, the preparation process of the hydrochloric acid and the mixed liquor of titanium source is as follows：It is 36~38% by mass concentration
Concentrated hydrochloric acid is 1 according to volume ratio with deionized water:1 mixing, is then added titanium source, the volume ratio of the titanium source and the concentrated hydrochloric acid
It is 1:25~1:60 stir evenly the mixed liquor up to the hydrochloric acid and titanium source.
Further, the titanium source is any one in butyl titanate, tetraethyl titanate, isopropyl titanate, titanium tetrachloride.
Further, the calcination processing in the step (1.5) specifically includes：Under inert gas protection, with 3~10
DEG C/heating rate of min is warming up to 250~300 DEG C, and 10~15min of constant temperature at a temperature of 250~300 DEG C, then with 3
The heating rate of~10 DEG C/min is warming up to 400~600 DEG C, and calcining at constant temperature 1~3 hour at a temperature of 400~600 DEG C,
Then control cooling.
Further, in step (2), the preparation process of the silver sol includes：By ethylene glycol and deionized water according to body
Product ratio 1:1 uniformly mixing, polyvinylpyrrolidone-K30 and sodium borohydride are then separately added into mixed solution, is stirred
After 1~2min, silver nitrate is added while stirring and is reacted, the silver nitrate:Polyvinylpyrrolidone-K30:Sodium borohydride
Molar ratio is 1:(0.01~0.03):(0.5~1), up to the silver sol after reaction.
Further, in step (3), the improvement cross-linking method specifically includes following steps：
(3.1) by 20~30mg bovine serum albumins be dissolved in 1.0mL pH value be 6.8 phosphate buffer in, then to
The glutaraldehyde of 80~150 μ L is added in the phosphate buffer, and bovine serum albumin-glutaraldehyde cross-linking agent solution is made, it will
The bovine serum albumin-glutaraldehyde cross-linking agent solution preserves at a temperature of 4 DEG C；
(3.2) 1~3mg glucose oxidases are dissolved in the phosphate buffer that 100 μ L pH value are 6.8, Portugal is made
Grape oxidase solution preserves the glucose oxidase solution at a temperature of 4 DEG C；
(3.3) the nano titania column array of nanometer modified by silver is immersed in phosphate buffer, so that described
The nano titania column array of nanometer modified by silver has the surface of moistening；
(3.4) 10~30 μ L bovine serum albumins-glutaraldehyde cross-linking agent solution is taken to be applied to the two of the nanometer modified by silver of moistening
Then TiOx nano column array surface stands 5~20min to carry out adsorption, is made and has adsorbed ox naturally at room temperature
The nano titania column array of the nanometer modified by silver of haemocyanin-glutaraldehyde cross-linking agent；
(3.5) it takes 10~30 μ L glucoses oxidation enzyme solutions to be applied to and has adsorbed bovine serum albumin-glutaraldehyde cross-linking agent
The surface of the nano titania column array of nanometer modified by silver at 4 DEG C stands 8~24 hours by cross-linking reaction by glucose
Oxidizing ferment is fixed, and is then soaked in phosphate buffer to remove loose glucose oxidase, is finally dried i.e.
Obtain the nano titania column array enzyme electrode of the nanometer modified by silver.
The second aspect of the present invention provides the nano titania column array enzyme electrode of a kind of nanometer of modified by silver, the nano silver
The nano titania column array enzyme electrode of modification is to be prepared using any one method in first aspect present invention,
In, the height of nano titania column is 1~3 μm, and the width of nano titania column is 50~150nm.
Further, the crystal form of the nano titania column array is rutile crystal type.
The third aspect of the present invention provides the nano titania column array enzyme of the nanometer modified by silver in above-mentioned second aspect
Application of the electrode in glucose biological sensor or enzyme thermistor devices.
The nano titania column array enzyme electrode of a kind of nanometer of modified by silver and its preparation method and application of the present invention, tool
It has the advantages that：
The present invention constructs TiO on carbon paper first using carbon paper as base material2Nano column array, then in TiO2Nanometer
The surface finish nano Argent grain of column array, obtains the TiO of nanometer modified by silver2Nano column array, then in this nanometer of modified by silver
TiO2Fixing glucose oxidase is adsorbed in the surface of nano column array using cross-linking method is improved, to obtain nanometer modified by silver
TiO2Nano column array enzyme electrode, preparation process is simple, both by means of the big specific surface of carbon paper to grow more TiO2Nano-pillar
Array, and TiO is utilized2The big specific surface area of nano column array, high electronics conduction efficiency and be conducive to electrolyte solution
To the higher electro catalytic activity of characteristic and nano-Ag particles of the diffusion of electrode surface, to substantially increase prepared enzyme electricity
The electro catalytic activity of pole.
The present invention nanometer modified by silver nano titania column array enzyme electrode manufacturing cost it is low, be easy to extensive life
Production, can be advantageously applied in glucose biological sensor and enzyme thermistor devices.
Description of the drawings
It, below will be to required in embodiment or description of the prior art in order to illustrate more clearly of technical scheme of the present invention
The attached drawing used is briefly described.It should be evident that drawings in the following description are only some embodiments of the invention, it is right
For those of ordinary skill in the art, without creative efforts, it can also be obtained according to these attached drawings
Its attached drawing.
Fig. 1 is the preparation method of the nano titania column array enzyme electrode of a kind of nanometer of modified by silver provided by the invention
Process flow chart, wherein A is carbon paper, and B is absorption TiO2The carbon paper of nano particle, C make a living with unformed TiO2Nano-pillar battle array
The carbon paper of row, D are nano titania column array/carbon paper, and E is the nano titania column array of nanometer modified by silver；
Fig. 2 is the SEM photograph of nano titania column array/carbon paper provided by the invention, wherein (2a) is front elevation,
(2b) is sectional view；
Fig. 3 is the XRD diagram of nano titania column array/carbon paper provided by the invention；
Fig. 4 is the SEM photograph of nano silver provided by the invention/nano titania column array/carbon paper；
Fig. 5 is to H2O2The current versus time curve of response, wherein (5a) is carbon paper electrode, and (5b) is nano titania
Column array/carbon paper electrode, (5c) are nano silver/nano titania column array/carbon paper electrodes；
Fig. 6 is carbon paper electrode (a), nano titania column array/carbon paper electrode (b) and nano silver/nano titania
The response current of column array/carbon paper electrode (c) is with H2O2The linear fit curve of concentration variation relation；
Fig. 7 is the current versus time curve to glucose responding, wherein (7a) is glucose oxidase/carbon paper enzyme electrode,
(7b) is glucose oxidase/nano titania column array/carbon paper enzyme electrode, and (7c) is glucose oxidase/nano silver/bis-
TiOx nano column array/carbon paper enzyme electrode；
Fig. 8 is glucose oxidase/carbon paper enzyme electrode (a), glucose oxidase/nano titania column array/carbon paper
The response current of enzyme electrode (b) and glucose oxidase/nano silver/nano titania column array/carbon paper enzyme electrode (c) is with Portugal
The linear fit curve of grape sugar concentration variation relation.
Specific implementation mode
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation describes, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, those of ordinary skill in the art obtained without making creative work it is all its
His embodiment, shall fall within the protection scope of the present invention.
Embodiment 1
The present embodiment provides the preparation method of the nano titania column array enzyme electrode of a kind of nanometer of modified by silver, such as Fig. 1
Shown, this method is as follows：
(1) using carbon paper as base material, TiO is grown on the carbon paper2Nano column array
It is by carbon paper cutting growth × width x thickness：The small pieces of 50mm × 20mm × 0.2mm use acetone and deionized water successively
Then supersound washing 10min is dried for standby at 80 DEG C, as shown in A in Fig. 1.
It takes 20mL butyl titanates in 200mL beakers, is slowly added to 100mL absolute ethyl alcohols while stirring, is then added dropwise
The glacial acetic acid of 0.2mL is stirred continuously to reaction solution is creamy white at room temperature, TiO is prepared2Colloidal solution.
By treated, spare carbon paper is immersed in above-mentioned TiO25min in colloidal solution, taking-up are dried at 80 DEG C, then
Cooled to room temperature after processing 20min, is prepared absorption TiO at 350 DEG C2The carbon paper of nano particle, such as B institutes in Fig. 1
Show.
Take 18mL H2The concentrated hydrochloric acid that 18mL mass concentrations are 36%~38% is added in 50mL beakers in O, stirs 5min,
Then the butyl titanate of 0.54mL is added into the hydrochloric acid solution, quickly stirs 10min, obtains the mixing of hydrochloric acid and butyl titanate
Liquid.
TiO will be adsorbed2The carbon paper of nano particle is put into the lining teflon reaction kettle of 50mL, then to the reaction
The mixed liquor of above-mentioned hydrochloric acid and butyl titanate is added in kettle, is reacted 10 hours at 180 DEG C.After reaction, it naturally cools to
Room temperature is respectively formed one layer of whiteness in the tow sides of carbon paper, and gained sample is rinsed with deionized water and removes material table repeatedly
The remaining reaction solution in face, dries, and growth, which is prepared, unformed TiO2The carbon paper of nano column array, as shown in C in Fig. 1.
Growth after above-mentioned dry there is into unformed TiO2The carbon paper of nano column array is placed in the program-controlled high temperature furnace of integration
Middle carry out calcination processing is warming up to 300 DEG C, and in the environment of 300 DEG C with the heating rate of 5 DEG C/min under nitrogen protection
Constant temperature 10min, then 500 DEG C are warming up to the heating rate of 5 DEG C/min, it then calcines 1.5 hours, forges in the environment of 500 DEG C
Cooling is controlled after burning, terminates program, the TiO after calcination processing2Nano column array from it is unformed be converted into more added with
The regular crystal phase of sequence has the TiO of crystal phase to get growth2The carbon paper of nano column array, hereinafter referred to as nano titania
Column array/carbon paper, as shown in D in Fig. 1.
The SEM photograph of the nano titania column array/carbon paper is as shown in Figure 2, wherein (2a) is its front elevation, (2b)
For its sectional view.By figure (2a) it is found that the surface of carbon paper surface and carbon paper inner void is all equal by nano titania column array
It covers evenly, nano titania column is in four prism type, and cylinder is smooth, and width is about 50~150nm, each titanium dioxide
Titanium nano-pillar is formed by multiple tiny nano titania column high-sequential close-packed arrays.By figure (2b) it is found that titanium dioxide
The height of nano-pillar is about 1~3 μm.
It is the XRD diagram of nano titania column array/carbon paper shown in Fig. 3, by the figure and Rutile Type TiO2Standard card
Piece (JCPDS No.21-1276) is compared with the standard card (JCPDS No.26-1076) of carbon pair it is found that the nano titania
The diffraction maximum of column and Rutile Type TiO2Standard card it is consistent, there is not apparent miscellaneous peak, the surface present invention is in carbon paper
The nano titania column of upper growth is rutile crystal type.
(2) preparation of the nano titania column array of nanometer modified by silver
Take 15mL H2O and 15mL ethylene glycol is mixed in the beaker of 50mL, is stirred evenly, and 1.2g polyethylene is then added
Pyrrolidones-K30, stirs and evenly mixs, and 0.0296g sodium borohydrides are added, and stirs 1~2min, and 0.1784g nitric acid is added while stirring
Silver, reaction solution become dark brown to get silver sol.
The nano titania column array/carbon paper prepared in step (1) is immersed in above-mentioned auburn silver sol, in
After constant temperature impregnates 10 hours at 40 DEG C, sample is taken out, is rinsed, is dried to get the dioxy of nanometer modified by silver repeatedly with deionized water
Change titanium nano column array, hereinafter referred to as nano silver/nano titania column array/carbon paper, as shown in E in Fig. 1.
Fig. 4 is the SEM photograph of the nano silver/nano titania column array/carbon paper, by figure it is found that silver nano-grain
It is uniformly deposited on the surface of nano titania column, the grain size of single Ag particles is about 10nm, there are a small amount of Ag particles in dioxy
The surface for changing titanium nano-pillar is reunited, and the particle of 30~40nm is gathered into.
(3) preparation of the nano titania column array enzyme electrode of nanometer modified by silver
Glucose oxidase is fixed on nano silver/nano titania column prepared by step (2) using cross-linking method is improved
Array/carbon paper surface, forms the nano titania column array enzyme electrode of nanometer modified by silver, and hereinafter referred to as grape is glycoxidative
Enzyme/nano silver/nano titania column array/carbon paper enzyme electrode, specific preparation process are as follows：
It takes 25mg bovine serum albumins in small centrifuge tube, the phosphate buffer that 1mL pH value is 6.8 is added, oscillation is mixed
It is even, the glutaraldehyde of 100 μ L is then added, is uniformly mixed, bovine serum albumin-glutaraldehyde cross-linking agent solution is made, and at 4 DEG C
Keep spare.
The glucose oxidase of 2.5mg is weighed in the small centrifuge tube of 1.5mL, the phosphoric acid that 100 μ L pH value are 6.8 is added
Salt buffer vibrates mixing, glucose oxidase solution is made, and saved backup at a temperature of 4 DEG C.
Nano silver/nano titania column array/carbon paper prepared by step (2) is dipped in phosphate buffer, is allowed to
Surface wettability.
20 μ L bovine serum albumins-glutaraldehyde cross-linking agent solution is taken to be applied to nano silver/nano titania column battle array of moistening
Then row/carbon paper surface stand 15min naturally at room temperature, the nanometer for having adsorbed bovine serum albumin-glutaraldehyde cross-linking agent is made
Silver/titanium dioxide nano column array/carbon paper；
20 μ L glucoses oxidation enzyme solutions are taken to be applied to nano silver/bis- for having adsorbed bovine serum albumin-glutaraldehyde cross-linking agent
The surface of TiOx nano column array/carbon paper stands 12 hours at 4 DEG C and carries out cross-linking reaction.Due to two aldehyde of glutaraldehyde
Base is reacted with the amino in bovine serum albumin and glucose oxidase molecules respectively, and therefore, glucose oxidase passes through glutaraldehyde
With bovine serum albumin be crosslinked, then by bovine serum albumin nano silver/nano titania column array/carbon paper surface suction
It is attached so that glucose oxidase is fixed on outer surface.It is soaked in after cross-linking reaction in the phosphate buffer that pH value is 6.8 to remove
Loose glucose oxidase is removed, is finally dried up to glucose oxidase/nano silver/nano titania column battle array
Row/carbon paper enzyme electrode.
Comparative example 1
Using the method for embodiment 1, glucose oxidase is directly anchored to pure carbon paper surface, it is glycoxidative that grape is made
Enzyme/carbon paper enzyme electrode.
Comparative example 2
Using the method for embodiment 1, nano titania column array/carbon paper is prepared, and glucose oxidase is fixed on
Prepared nano titania column array/carbon paper surface forms glucose oxidase/nano titania column array/carbon paper
Enzyme electrode.
Embodiment 2
The present embodiment respectively with pure carbon paper, comparative example 2 prepare nano titania column array/carbon paper and embodiment 1 in
The nano silver of preparation/nano titania column array/carbon paper is as working electrode, using platinum plate electrode as to electrode, with saturation
Calomel electrode is as reference electrode, under the operating potential of -0.6V, is electrolysed by work of the phosphate buffer that pH value is 6.8
Matter solution carries out current versus time curve test, and three is to H for detection2O2Electrocatalysis characteristic, wherein H2O2Glucose oxidase
It is catalyzed the product of reaction.
Specific detection process is that a H is added per 50s2O2Solution, since 100s, each increased H2O2Concentration increases
(0.1mmol ﹒ L are indicated for 0.1mM-1, same as below)；Since 600s, each increased H2O2Concentration increases to 0.2mM；From
850s starts, each increased H2O2Concentration increases to 0.5mM, until test terminates, testing result is as shown in Figure 5 and Figure 6.Fig. 5
For to H2O2The current versus time curve of response, wherein (5a) is pure carbon paper electrode, and (5b) is nano titania column array/carbon
Paper electrode, (5c) are nano silver/nano titania column array/carbon paper electrode；Fig. 6 is carbon paper electrode (a), nano titania
The response current of column array/carbon paper electrode (b) and nano silver/nano titania column array/carbon paper electrode (c) is with H2O2Concentration
The linear fit curve of variation relation.
By testing result it is found that carbon paper electrode is to H2O2The linear concentration of response ranging from 0~7mM, nano titania column
Array/carbon paper electrode is to H2O2The linear concentration of response ranging from 0~3mM, nano silver/nano titania column array/carbon paper electricity
Extremely to H2O2The linear concentration of response ranging from 0~5mM；Carbon paper electrode is to H2O2Responsive current density be 0.0325mA ﹒ mM-1﹒
cm-2, nano titania column array/carbon paper electrode is to H2O2Responsive current density be 1.138mA ﹒ mM-1﹒ cm-2, nano silver/
Nano titania column array/carbon paper electrode is to H2O2Responsive current density be 1.62mA ﹒ mM-1﹒ cm-2, it is seen then that nano silver/
Nano titania column array/carbon paper electrode is to H2O2Responsive current density be 50 times of carbon paper electrode, and be that titanium dioxide is received
1.42 times of rice column array/carbon paper electrode.
Therefore, compared with carbon paper electrode and nano titania column array/carbon paper electrode, nano silver/nano titania
Column array/carbon paper electrode is to H2O2With better electrocatalysis characteristic.
Embodiment 3
The present embodiment is respectively with the glucose oxidase/carbon paper prepared in comparative example 1, the glucose prepared in comparative example 2
Glucose oxidase/nanometer the silver/titanium dioxide prepared in oxidizing ferment/nano titania column array/carbon paper and embodiment 1 is received
Rice column array/carbon paper is working electrode, using platinum plate electrode as to electrode, using saturated calomel electrode as reference electrode, with pH
The phosphate buffer that value is 6.8 is working electrolyte solution, carries out current versus time curve test, three is to glucose for detection
Electrocatalysis characteristic, wherein test glucose oxidase/carbon paper enzyme electrode when select -0.5V be used as operating potential, test grape
Select -0.6V as operating potential when carbohydrate oxidase/nano silver/nano titania column array/carbon paper enzyme electrode.
Specific detection process is that a glucose solution is added per 50s, since 100s, each increased glucose
Concentration increases to 0.05mM；Since 400s, each increased concentration of glucose increases to 0.1mM, until test terminates, detection knot
Fruit is as shown in Figure 7 and Figure 8.Fig. 7 is the current versus time curve to glucose responding, wherein (7a) is glucose oxidase/carbon
Paper enzyme electrode, (7b) be glucose oxidase/nano titania column array/carbon paper enzyme electrode, (7c) be glucose oxidase/
Nano silver/nano titania column array/carbon paper enzyme electrode；Fig. 8 is glucose oxidase/carbon paper enzyme electrode (a), grape glycosyloxy
Change enzyme/nano titania column array/carbon paper enzyme electrode (b) and glucose oxidase/nano silver/nano titania column battle array
The response current of row/carbon paper enzyme electrode (c) with concentration of glucose variation relation linear fit curve.
By testing result it is found that glucose oxidase/carbon paper enzyme electrode is to the responsive current density of glucose
0.0276mA ﹒ mM-1﹒ cm-2, response electricity of the glucose oxidase/nano titania column array/carbon paper enzyme electrode to glucose
Current density is 0.13mA ﹒ mM-1﹒ cm-2, glucose oxidase/nano silver/nano titania column array/carbon paper enzyme electrode is to Portugal
The responsive current density of grape sugar is 0.182mA ﹒ mM-1﹒ cm-2, it is seen then that glucose oxidase/nano silver/nano titania column
Array/carbon paper enzyme electrode is Portugal to 6.6 times that the responsive current density of glucose is glucose oxidase/carbon paper enzyme electrode
1.4 times of grape carbohydrate oxidase/nano titania column array/carbon paper enzyme electrode.
Therefore, with glucose oxidase/carbon paper enzyme electrode and glucose oxidase/nano titania column array/carbon paper
Enzyme electrode is compared, and glucose oxidase/nano silver/nano titania column array/carbon paper enzyme electrode has glucose more preferable
Electrocatalysis characteristic.
Glucose oxidase/nano silver/nano titania column array/carbon paper enzyme electrode is to the linear dense of glucose responding
Spend ranging from 0~0.9mM, linearly dependent coefficient R2=0.9996, detection is limited to 1.6 μM (signal-to-noise ratio 3).
To sum up, the present invention constructs TiO on carbon paper first using carbon paper as base material2Then nano column array exists
TiO2The surface finish nano Argent grain of nano column array, obtains the TiO of nanometer modified by silver2Nano column array, then in the nano silver
The TiO of modification2Fixing glucose oxidase is adsorbed in the surface of nano column array using cross-linking method is improved, and is repaiied to obtain nano silver
The TiO of decorations2Nano column array enzyme electrode, preparation process is simple, both by means of the big specific surface of carbon paper to grow more TiO2It receives
Rice column array, and TiO is utilized2The big specific surface area of nano column array, high electronics conduction efficiency and be conducive to electrolyte
The higher electro catalytic activity of characteristic and nano-Ag particles from solution to the diffusion of electrode surface so that prepared nano silver
The nano titania column array enzyme electrode of modification has better electro catalytic activity.The titanium dioxide of the nanometer modified by silver of the present invention
The manufacturing cost of titanium nano column array enzyme electrode is low, is easy to mass produce, and can be advantageously applied to glucose biological sensor
And in enzyme thermistor devices.
Above description has fully disclosed the specific implementation mode of the present invention.It should be pointed out that being familiar with the field
Technical staff is to any change for being done of specific implementation mode of the present invention all without departing from the range of claims of the present invention.
Correspondingly, the scope of the claims of the invention is also not limited only to previous embodiment.
Claims (10)
1. the preparation method of the nano titania column array enzyme electrode of a kind of nanometer of modified by silver, which is characterized in that the method
Include the following steps：
(1) using carbon paper as base material, TiO is grown on the carbon paper2Nano column array；
(2) growth in step (1) there is into TiO2The carbon paper of nano column array is immersed in silver sol 3-15 hours, is then taken out, and is used
The nano titania column array of nanometer modified by silver is drying to obtain after deionized water cleaning；
(3) with bovine serum albumin-glutaraldehyde as cross linker, glucose oxidase is fixed on step (2) using cross-linking method is improved
Described in nanometer modified by silver nano titania column array surface, that is, form the nano titania of the nanometer modified by silver
Column array enzyme electrode.
2. the preparation method of the nano titania column array enzyme electrode of a kind of nanometer of modified by silver according to claim 1,
It is characterized in that, in step (1), it is described using carbon paper as base material, TiO is grown on carbon paper2Nano column array, it is specific to wrap
Include following steps：
(1.1) carbon paper of certain size size is taken, it is spare after cleaning, drying；
(1.2) it is 1 according to volume ratio by titanium source and absolute ethyl alcohol:4~1:20 mixing, then to the mixed solution and dripping ice
The volume ratio of acetic acid, the glacial acetic acid and titanium source is 1:50~1:250, it is stirred to react to obtain TiO at room temperature2Colloidal solution；
(1.3) TiO being immersed in the carbon paper being dried for standby in step (1.1) in step (1.2)2In colloidal solution, then take out
Drying handles 10~30min postcoolings to room temperature at 300~400 DEG C, obtains absorption TiO2The carbon paper of nano particle；
(1.4) by the absorption TiO2The carbon paper of nano particle is put into hydrochloric acid and the mixed liquor of titanium source, anti-at 150~200 DEG C
It answers 4~20 hours, product obtained by the reaction is washed off dry after the reaction solution of remained on surface with deionized water, and obtaining growth has
Unformed TiO2The carbon paper of nano column array；
(1.5) product after will be dry in step (1.4), which is placed in the high temperature furnace of inert gas atmosphere, carries out calcination processing, calcines
Temperature is 400~600 DEG C, and calcination time is 1~3 hour, and heating rate is 3~10 DEG C/min, and then control cools down up to raw
TiO with crystal phase2The carbon paper of nano column array.
3. the preparation method of the nano titania column array enzyme electrode of a kind of nanometer of modified by silver according to claim 2,
It is characterized in that, the preparation process of the hydrochloric acid and the mixed liquor of titanium source is as follows：The concentrated hydrochloric acid for being 36~38% by mass concentration
With deionized water according to volume ratio be 1:1 mixing, is then added titanium source, and the volume ratio of the titanium source and the concentrated hydrochloric acid is 1:25
~1:60, stir and evenly mix the mixed liquor up to the hydrochloric acid and titanium source.
4. the preparation side of the nano titania column array enzyme electrode of a kind of nanometer of modified by silver according to claim 2 or 3
Method, which is characterized in that the titanium source is any one in butyl titanate, tetraethyl titanate, isopropyl titanate, titanium tetrachloride.
5. the preparation method of the nano titania column array enzyme electrode of a kind of nanometer of modified by silver according to claim 2,
It is characterized in that, the calcination processing in the step (1.5) specifically includes：
Under inert gas protection, 250~300 DEG C are warming up to the heating rate of 3~10 DEG C/min, and at 250~300 DEG C
At a temperature of 10~15min of constant temperature, be then warming up to 400~600 DEG C with the heating rate of 3~10 DEG C/min, and 400~600
Calcining at constant temperature 1~3 hour at a temperature of DEG C, then control cooling.
6. the preparation method of the nano titania column array enzyme electrode of a kind of nanometer of modified by silver according to claim 1,
It is characterized in that, in step (2), the preparation process of the silver sol includes：
By ethylene glycol and deionized water according to volume ratio 1:1 uniformly mixing, is then separately added into poly- second into mixed solution
Alkene pyrrolidone-K30 and sodium borohydride after stirring 1~2min, are added silver nitrate and are reacted while stirring, the silver nitrate:
Polyvinylpyrrolidone-K30:The molar ratio of sodium borohydride is 1:(0.01~0.03):(0.5~1), up to the silver after reaction
Colloidal sol.
7. the preparation method of the nano titania column array enzyme electrode of a kind of nanometer of modified by silver according to claim 1,
It is characterized in that, in step (3), the improvement cross-linking method specifically includes following steps：
(3.1) 20~30mg bovine serum albumins are dissolved in the phosphate buffer that 1.0mL pH value is 6.8, then to described
The glutaraldehyde of 80~150 μ L is added in phosphate buffer, and bovine serum albumin-glutaraldehyde cross-linking agent solution is made, it will be described
Bovine serum albumin-glutaraldehyde cross-linking agent solution preserves at a temperature of 4 DEG C；
(3.2) 1~3mg glucose oxidases are dissolved in the phosphate buffer that 100 μ L pH value are 6.8, glucose is made
Enzyme solutions are aoxidized, the glucose oxidase solution is preserved at a temperature of 4 DEG C；
(3.3) the nano titania column array of nanometer modified by silver is immersed in phosphate buffer, so that the nanometer
The nano titania column array of modified by silver has the surface of moistening；
(3.4) 10~30 μ L bovine serum albumins-glutaraldehyde cross-linking agent solution is taken to be applied to the titanium dioxide of the nanometer modified by silver of moistening
Then titanium nano column array surface stands 5~20min to carry out adsorption, is made and has adsorbed cow's serum naturally at room temperature
The nano titania column array of the nanometer modified by silver of albumen-glutaraldehyde cross-linking agent；
(3.5) 10~30 μ L glucoses oxidation enzyme solutions is taken to be applied to the nanometer for having adsorbed bovine serum albumin-glutaraldehyde cross-linking agent
The surface of the nano titania column array of modified by silver, stood at 4 DEG C 8~24 hours it is by cross-linking reaction that grape is glycoxidative
Enzyme is fixed, and is then soaked in phosphate buffer to remove loose glucose oxidase, is finally dried up to institute
State the nano titania column array enzyme electrode of nanometer modified by silver.
8. the nano titania column array enzyme electrode of a kind of nanometer of modified by silver, which is characterized in that using in claim 1~7
The preparation method of the nano titania column array enzyme electrode of a kind of nanometer of modified by silver described in any one is made, the dioxy
The height for changing titanium nano-pillar is 1~3 μm, and the width of nano titania column is 50~150nm.
9. the nano titania column array enzyme electrode of a kind of nanometer of modified by silver according to claim 8, which is characterized in that
The crystal form of the nano titania column array is rutile crystal type.
10. the nano titania column array enzyme electrode of a kind of nanometer of modified by silver described in claim 8 or 9 is given birth in glucose
Application in object sensor or enzyme thermistor devices.
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| CN109794237B (en) * | 2019-01-10 | 2021-08-13 | 北京工业大学 | Preparation method of silver-loaded titanium dioxide nanorod array photocatalyst |
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| CN109975375B (en) * | 2019-04-11 | 2021-07-02 | 福建师范大学 | Zearalenone detection method based on signal enhancement type polymer functionalized rutile phase titanium dioxide mesocrystal |
| CN110146570A (en) * | 2019-05-09 | 2019-08-20 | 清华-伯克利深圳学院筹备办公室 | Enzyme electrode and preparation method thereof, enzyme sensor, monitoring device and therapeutic equipment |
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| CN114311865B (en) * | 2021-12-15 | 2023-08-15 | 佛山(华南)新材料研究院 | Laminated TiO 2 Preparation method of heat conducting material |
| CN116046874A (en) * | 2023-02-15 | 2023-05-02 | 重庆联芯致康生物科技有限公司 | Dynamic alcohol sensor structure and preparation method |
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