CN105973962A - Preparation method of glucose sensor based on graphene nano wall - Google Patents
Preparation method of glucose sensor based on graphene nano wall Download PDFInfo
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- CN105973962A CN105973962A CN201610261475.5A CN201610261475A CN105973962A CN 105973962 A CN105973962 A CN 105973962A CN 201610261475 A CN201610261475 A CN 201610261475A CN 105973962 A CN105973962 A CN 105973962A
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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
Abstract
The invention provides a preparation method of a glucose sensor; a graphene nano wall is used as a loading skeleton, through research and technology improvement of growth conditions of the key sensor substrate-three-dimensional graphene wall, sensor preparation and glucose oxidase function technology are optimized, and the novel three-dimensional graphene glucose sensor can be prepared; the sensor has high electrochemical response sensitivity on glucose, and shows relatively good linear responsiveness and excellent glucose sensing specificity. The production process of the product is simple, low in cost, and suitable for large-scale industrialized mass production.
Description
Technical field
The present invention relates to the preparation method of a kind of glucose sensor.
Background technology
Diabetes are one group
The metabolic disease of feature.Diabetes have become a transworld Community health problem, have become as continue swollen
The chronic disease of serious threat human health after tumor, cardiovascular and cerebrovascular disease.Long-term diabetes can cause many
Complication, such as heart disease, renal failure, blind and peripheral neuropathy etc., and strict, personalized blood glucose control
System can reduce the appearance of complication significantly.In recent years, glucose biological sensor is because of easy to carry, sensitive
The features such as degree is high, selectivity is good, detection speed is fast, simple to operate receive much concern.Building by biosensor can
It is divided into and has enzyme and enzyme-free glucose electrochemical sensor, wherein have enzyme sensor because its processing technology is simple, sensing
High specificity and extensively researched and developed.
Graphene, a kind of by the former molecular New Two Dimensional c-based nanomaterial of monolayer carbon, lead at bio electrochemistry
Research in territory is of increasing concern, and its excellent electrocatalysis characteristic becomes prepares enzyme electrochemical biosensor
One of preferable electrode material of device, its special molecular structure makes it can be effectively facilitated electric transmission, promotes
The response signal of biosensor and sensitivity.Additionally, its good biocompatibility can keep sensor institute
The biological activity of load enzyme, improves the stability of sensor.Recently, graphene nano wall, a kind of by Graphene
Nanometer sheet vertical distribution graphene three-dimensional structure on base material, has the richest compared with two-dimensional graphene
Rich morphosis and huge specific surface area, impart more preferable draftability simultaneously, have started to gradually be applied to
The research and development of sensor, as to gas, the monitoring of temperature.
Application No. 201410235183.5 patent document discloses a kind of electrochemical glucose based on Graphene
Sensor and preparation method thereof, can measure glucose content, but the preparation of this sensor needs to carry out metal to be received
Rice grain is modified so that product preparation process is complex.This patent uses three-dimensional grapheme nanometer simultaneously
Pore structure, after modifying through metallic particles, its electrical conductivity, biocompatibility can be affected.
Summary of the invention
For the deficiencies in the prior art, it is an object of the invention to provide a kind of graphene nano wall glucose sensing
The preparation method of device, prepared product electrochemical workstation test has shown height linear response,
Higher specificity and sensitivity, have higher electrical conductivity, the conductive network structure of more horn of plenty and material
Bio-compatibility is good.
It is an object of the invention to be realized by following measures:
The preparation method of a kind of glucose sensor, uses graphene nano wall as load skeleton, described graphite
Alkene nm wall using plasma strengthens chemical vapour deposition technique (PECVD) to be prepared, Copper Foil successively carries out heating up,
Annealing, growth step；Wherein, Elevated Temperature Conditions is 30～90min to be warming up to 500～800 DEG C, hydrogen=5～
30sccm；Annealing conditions is 30～90min, maintains 600～800 DEG C, hydrogen=5～15sccm；Growth bar
Part is 600～800 DEG C and maintains 60～120min, and methane: hydrogen is 4: 8sccm, radio frequency is 200～500mw；
Closing radio frequency and methane, hydrogen is set to 2～20sccm, and taking out growth after cooling has the Copper Foil of Graphene wall；
After graphene nano wall is transferred to polyethylene terephthalate (PET), use silk screen print method
After printing three electrodes, electrode is carried out surface-functionalized being allowed to and loads glucoseoxidase.
In order to improve susceptiveness and the specificity of sensor further, above-mentioned surface functionalization step is: a) will
The glucose oxidase solution of 100-800IU/ml loads to three-dimensional grapheme nm wall, dries；B) will
0.3-3.5wt% chitosan solution drips and is layed onto three-dimensional grapheme/glucoseoxidase surface；C) by 0.25-2wt%'s
Glutaraldehyde is fixed to three-dimensional grapheme wall/enzyme/chitosan surface, dries；D) PBS solution cleaning electrode is used
And preserve.Glucoseoxidase is directly fixed to by the present invention by the chitosan-glutaraldehyde cross-linking absorption optimized
Nm wall surface i.e. prepares electrode.
Above-mentioned PBS solution concentration is 0.1M, pH=7.0, cleans and storage temperature is 2-8 DEG C.
Above-mentioned employing silk screen print method printed electrode step is: Ag/AgCl ink a) is printed on PET work
For reference electrode, toast 5-25 minute 60-100 DEG C of scope, b) carbon printing ink is printed on PET as work
Make electrode and to electrode, toast 15-30 minute 80-120 DEG C of scope, c) print one again at working electrode surface
Layer insulation paste, toasts in the range of 50-90 DEG C 5-20 minute, d) three electrodes connects conduction filamentary silver respectively and draw
Go out circuit.
Specifically, the preparation method of the graphene nano wall glucose sensor of the present invention, comprise the following steps:
(1) preparation of Graphene wall
Use PECVD, the Copper Foil that thickness is 1 μm～100 μm is lain in a horizontal plane in the glass tubing of growth furnace
In, successively through heating up, anneal, growing three steps；Wherein, temperature-rise period is 30～90min to be warming up to
500～800 DEG C, hydrogen=5～30sccm；Annealing process is 30～90min, maintains 600～800 DEG C, hydrogen
Gas=5～15sccm；Growth course is 60～120min, maintains 600～800 DEG C, methane: hydrogen=4: 8sccm,
Radio frequency=200～500mw；Growth closes radio frequency and methane after terminating, and hydrogen arranges 2～20sccm, opens
Fan fast cooling, the most gently taking out growth has the Copper Foil of Graphene wall；
(2) transfer of Graphene wall
A) compound concentration is the Fe (NO of 0.1M～0.5M3)3Copper etching liquid, b) is cut into Copper Foil a size of
0.2cm*0.2cm～3cm*3cm fritter not etc. inserts solution surface, has the one of Graphene wall to face with growth
On, overnight fully to etch Copper Foil under room temperature condition, c) with plasma treated size be 0.2cm*0.2cm～
Floating Graphene wall is picked up by the PET film that 3cm*3cm does not waits, and is transferred in ultra-pure water by Graphene wall
30min*3～5 times, fully to remove the metal ions such as remaining copper, ferrum, d) pick up cleaned stone with PET
Ink alkene wall also puts into drying under conditions of 80～150 DEG C * 5～60min in baking oven, makes Graphene wall and PET
Secure fit；
(3) preparation of electrode
Use silk screen print method printed electrode, a) Ag/AgCl ink be printed on PET as reference electrode,
Toast 5-25 minute 60-100 DEG C of scope, b) carbon printing ink is printed on PET as working electrode with to electricity
Pole, toasts 15-30 minute 80-120 DEG C of scope, c) prints one layer of insulation paste again at working electrode surface,
Toast in the range of 50-90 DEG C 5-20 minute, d) three electrodes are connected respectively conduction filamentary silver and draw circuit；
(4) sensor electrode is surface-functionalized
A) configuration concentration is the glucose oxidase solution of 100-800IU/ml, is aoxidized by 2-20 μ L glucose
Enzymatic solution loads on three-dimensional grapheme nm wall, and room temperature is dried, and b) by 2-20 μ L, concentration is 0.3-3.5%
Chitosan solution drips and is layed onto three-dimensional grapheme/glucoseoxidase surface, is c) glutaraldehyde of 0.25-2% by concentration
Being fixed to three-dimensional grapheme wall/enzyme/chitosan surface, room temperature is dried a period of time, d) remnants on cleaning electrode
Glucoseoxidase also preserves；During cleaning and preserving, used PBS solution concentration is 0.1M,
PH=7.0, the temperature of process is 2-8 DEG C.
In order to improve the sensor sensitivity to glucose detection further, after prepared by electrode, surface-functionalized
Before, use oxygen plasma that graphene nano wall surface is modified；Described modification condition is: oxygen flow
For 5-30sccm, air pressure is that 0.1-1mbar processes 1-10min.
Beneficial effect
1. the present invention is by changing the research of the growth conditions of key sensor base material-three-dimensional grapheme wall and technology
Entering, use silk screen print method to print three electrodes, the chitosan-glutaraldehyde cross-linking absorption method by optimizing will simultaneously
Glucoseoxidase is directly fixed to nm wall surface and can be prepared by electrode.Electrode test shows: use the method
The electrode of preparation, has good susceptiveness and specificity.
2., after the present invention uses oxygen plasma that graphene nano wall surface is carried out moditied processing, carry further
The high sensor sensitivity to glucose detection.
3. the present invention prepares glucose sensor without carrying out metal nanoparticle modification, the most preferably avoids
Because metal nanoparticle modifies the impact bringing Graphene electrical conductivity, biocompatibility, and technique
Simply, with low cost, without accurately controlling technological parameter, be suitable to large-scale industrialization batch production.
4. the present invention prepares novel three-dimensional grapheme glucose sensor, and this sensor is for the electricity of glucose
Chemical response is sensitive, and embodies the linear response of height, higher specificity；This graphene nano wall
With low cost, product processes is simple, is suitable to large-scale industrialization batch production.
Accompanying drawing explanation
Fig. 1 is the preparation technology stream of a kind of three-dimensional grapheme nm wall glucose sensor involved in the present invention
Cheng Tu；
Fig. 2 is the three-dimensional grapheme nm wall high resolution scanning electromicroscopic photograph obtained by embodiment 1: A is low power
The surface texture of graphene nano wall under mirror；B is the surface texture of graphene nano wall under high power lens；C is high
The cross section structure of graphene nano wall under times mirror；D is the space structure schematic diagram of three-dimensional grapheme nm wall.
Fig. 3 is the three-dimensional grapheme nm wall glucose sensor of preparation in embodiment 1, successively containing Fructus Vitis viniferae
The phosphate buffer (PBS) of 0.1M, pH=7.0 of sugar (1mM) is circulated volt-ampere curve test.
In figure, test operation voltage is-1.0-1.0V, and sweep speed is 100mV/s.
Fig. 4 is the three-dimensional grapheme nm wall glucose sensor of preparation in embodiment 1, successively containing Fructus Vitis viniferae
The phosphate buffer (PBS) of 0.1M, pH=7.0 of sugar (1mM) carries out time current curve test.
In figure, test operation voltage is 0.3V vs.Ag/AgCl.
Fig. 5 is that the three-dimensional grapheme glucose sensor product introduction obtained by embodiment 1 has gone out highly linear sound
Ying Xing.In figure, test operation voltage is 0.3V vs.Ag/AgCl.
Fig. 6 is that the three-dimensional grapheme glucose sensor product introduction obtained by embodiment 1 has gone out the special of height
Property.In figure, concentration of glucose is 2mM, and uric acid concentration is 1mM, and vitamin A concentration is 0.2mM, surveys
Examination operation voltage is 0.3V vs.Ag/AgCl.
Fig. 7 is that three-dimensional grapheme scanning electron microscopic observation after oxygen plasma processes that embodiment 5 is grown finds
There is not significant change in graphene nano wall surface texture, and process group (B) with without oxygen plasma treatment group
(A) compare and loaded more glucoseoxidase/chitosan/glutaraldehyde cross-linking body.
Fig. 8 is the glucose sensing that the three-dimensional grapheme that embodiment 5 is grown prepares after oxygen plasma processes
Device product (blue) with embodied compared with oxygen plasma treatment group (red) higher glucose detection spirit
Quick property.In figure, concentration of glucose is 2mM, and test operation voltage is 0.3V vs.Ag/AgCl.
Detailed description of the invention
The present invention will be described with implementing row below in conjunction with the accompanying drawings.The description below embodiment carried out be in order to
It is easy to those skilled in the art understand and apply the invention.Person skilled in the art is obvious
Easily these embodiments can be made various amendment, and it is real that General Principle described herein is applied to other
Execute in example without through performing creative labour.Therefore, the invention is not restricted to embodiment here, this area skill
Art personnel should be at the protection model of the present invention according to the announcement of the present invention, the improvement making the present invention and amendment
Within enclosing.
Implement row 1
The present embodiment provides the preparation method of a kind of three-dimensional grapheme glucose sensor, the technological process of the method
Figure is as it is shown in figure 1, specifically comprise the following steps that
1. use PECVD growing three-dimensional Graphene: the Copper Foil of 20 μ m-thick is lain in a horizontal plane in growth furnace
In glass tubing, successively through heating up, anneal, growing three steps.Condition is respectively as follows: intensification: 30min liter
Warm to 500 DEG C, hydrogen=5,10,15,20,25,30sccm；Annealing: 30min, maintains 600 DEG C,
Hydrogen=5sccm；Growth: 60min, maintains 600 DEG C, methane: hydrogen=4: 8sccm, radio frequency=200mw；Raw
Length closes radio frequency and methane after terminating, and hydrogen arranges 2sccm, opens fan fast cooling, the most gently
Take out growth and have the Copper Foil of Graphene wall.
2. three-dimensional grapheme is transferred to PET base material: prepares Fe (NO3) the 3 bronze medal etching liquid of 0.1M, then will
Copper Foil is cut to the fritter of 0.5cm*0.5cm and inserts solution surface, has the one of Graphene wall to face up with growth,
Overnight fully to etch Copper Foil under room temperature condition.Then, (the hydrophilic of pet sheet face is exposed with plasma treated
Group, convenient with the laminating of Graphene wall) 1cm*1cm PET film floating Graphene wall is picked up, by stone
Ink alkene wall is transferred in ultra-pure water remove the metal ions such as the copper of remnants, ferrum with abundant 30min*3 time.Then,
PET picks up cleaned Graphene wall and puts in baking oven and to toast 5min under conditions of 80 DEG C, makes Graphene
Wall and PET secure fit.
3. use silk screen print method printed electrode: be first printed on as reference electrode on PET using Ag/AgCl ink,
70 DEG C are toasted 5 minutes；Then, carbon printing ink is printed on PET as working electrode with to electrode, 80 DEG C of bakings
Roasting 15 minutes；Printing one layer of insulation paste again at working electrode surface again, 70 DEG C are toasted 10 minutes；Finally will
Three electrodes connect conduction filamentary silver respectively and draw circuit.
4. three-dimensional graphene electrode is surface-functionalized: weighing 0.5g chitosan, to be dissolved in the acetic acid of 100ml 0.5% molten
In liquid, room temperature stirs 3h to prepare chitosan solution with 250 revs/min；Draw 3 μ L glucose oxidase
Solution (concentration is 100IU/mL) drop coating is on GNWs, and room temperature is dried；Take 3 μ L 0.5% chitosans molten
Drop is coated onto GNWs/ enzyme surface, and room temperature is dried；Take the glutaraldehyde drop coating of 3 μ L0.25% be layed onto GNWs/ enzyme/
Chitosan surface, room temperature dries 1h；Electrode is immersed in the 0.1M PBS of pH 7.0,4 DEG C of mistakes
Night is to remove glucoseoxidase remaining on electrode；4 DEG C are saved in the 0.1M PBS of pH7.0.
The most then on electrochemical workstation, the analytical performance of sensor is tested: 1) containing glucose (1mM)
0.1M, pH=7.0 phosphate buffer (PBS) in carry out the circulation of three-dimensional grapheme glucose sensor
Volt-ampere curve test (Fig. 3).Wherein, test voltage is-1.0-1.0v, and sweep speed is 100mv/s.2)
Three-dimensional stone is carried out in the phosphate buffer (PBS) of 0.1M, pH=7.0 containing glucose (1mM)
Time current curve test (Fig. 4) of ink alkene glucose sensor.Wherein, test voltage is 0.3V.3)
The concentration 0.5-0.9mM glucose that obtained product sensor is sequentially placed into the preparation of above-mentioned PBS is molten
Liquid carries out linear response test, records 60s current value size (Fig. 5) of reaction in each concentration.Its
In, test voltage is 0.3V.4) being 2mM at concentration of glucose, uric acid concentration is 1mM, vitamin A
Concentration be 0.1M, pH=7.0 of 0.2mM PBS in carry out product specificity test, wherein, test electricity
Pressure is 0.3V.
Implement row 2
The present embodiment provides the preparation method of a kind of three-dimensional grapheme glucose sensor, the technological process of the method
Figure is as it is shown in figure 1, specifically comprise the following steps that
1. use PECVD growing three-dimensional Graphene: the Copper Foil of 60 μ m-thick is lain in a horizontal plane in growth furnace
In glass tubing, successively through heating up, anneal, growing three steps.Condition is respectively as follows: intensification: 50min
It is warming up to 700 DEG C, hydrogen=25sccm；Annealing: 70min, maintains 750 DEG C, hydrogen=10sccm；Growth:
100min, maintains 800 DEG C, methane: hydrogen=9: 6sccm, radio frequency=400mw；Growth is closed after terminating and is penetrated
Frequency and methane, hydrogen arranges 10sccm, opens fan fast cooling, and the most gently taking out growth has stone
The Copper Foil of ink alkene wall.
2. three-dimensional grapheme is transferred to PET base material: prepares Fe (NO3) the 3 bronze medal etching liquid of 0.5M, then will
Copper Foil is cut to the fritter of 2.0cm*2.0cm and inserts solution surface, has the one of Graphene wall to face up with growth,
Overnight fully to etch Copper Foil under room temperature condition.Then, (the hydrophilic of pet sheet face is exposed with plasma treated
Group, convenient with the laminating of Graphene wall) 4cm*5cm PET film floating Graphene wall is picked up, by stone
Ink alkene wall is transferred in ultra-pure water remove the metal ions such as the copper of remnants, ferrum with abundant 30min*4 time.Then,
PET picks up cleaned Graphene wall and puts into drying under conditions of 100 DEG C of * 15min in baking oven, makes graphite
Alkene wall and PET secure fit.
3. use silk screen print method printed electrode: be first printed on as reference electrode on PET using Ag/AgCl ink,
70 DEG C are toasted 10 minutes；Then, carbon printing ink is printed on PET as working electrode with to electrode, 120 DEG C
Toast 15 minutes；Printing one layer of insulation paste again at working electrode surface again, 80 DEG C are toasted 10 minutes；?
After by three electrodes respectively connect conduction filamentary silver draw circuit.
4. three-dimensional graphene electrode is surface-functionalized: weighs 2.5g chitosan and is dissolved in the acetic acid of 100ml 2.5%
In solution, room temperature stirs 3h to prepare chitosan solution with 250 revs/min；Draw 20 μ L glucose
Oxidase solution (concentration is 500IU/mL) drop coating is on GNWs, and room temperature is dried；Take 20 μ L 2.5% shells
Polysaccharide solution drop coating is to GNWs/ enzyme surface, and room temperature is dried；The glutaraldehyde drop coating taking 20 μ L2.0% is layed onto GNWs/
Enzyme/chitosan surface, room temperature dries 1h；Electrode is immersed in the 0.1M PBS of pH 7.0,4 DEG C
Overnight to remove glucoseoxidase remaining on electrode；4 DEG C are saved in the 0.1M PBS of pH7.0.
Implement row 3
The present embodiment provides the preparation method of a kind of three-dimensional grapheme glucose sensor, the technological process of the method
Figure is as it is shown in figure 1, specifically comprise the following steps that
1. use PECVD growing three-dimensional Graphene: the Copper Foil of 50 μ m-thick is lain in a horizontal plane in growth furnace
In glass tubing, successively through heating up, anneal, growing three steps.Condition is respectively as follows: intensification: 45min liter
Warm to 650 DEG C, hydrogen=20sccm；Annealing: 60min, maintains 750 DEG C, hydrogen=5sccm；Growth: 90min,
Maintain 750 DEG C, methane: hydrogen=10: 8sccm, radio frequency=350mw；Growth closes radio frequency and methane after terminating,
Hydrogen arranges 8sccm, opens fan fast cooling, and the most gently taking out growth has the Copper Foil of Graphene wall.
2. three-dimensional grapheme is transferred to PET base material: prepares Fe (N03) the 3 bronze medal etching liquid of 0.4.M, then will
Copper Foil is cut to the fritter of 1.5cm*1.5cm and inserts solution surface, has the one of Graphene wall to face up with growth,
Overnight fully to etch Copper Foil under room temperature condition.Then, (the hydrophilic of pet sheet face is exposed with plasma treated
Group, convenient with the laminating of Graphene wall) 4cm*4cm PET film floating Graphene wall is picked up, by stone
Ink alkene wall is transferred in ultra-pure water remove the metal ions such as the copper of remnants, ferrum with abundant 30min*3 time.Then,
PET picks up cleaned Graphene wall and puts into drying under conditions of 100*10min in baking oven, makes Graphene
Wall and PET secure fit.
3. use silk screen print method printed electrode: be first printed on as reference electrode on PET using Ag/AgCl ink,
100 DEG C are toasted 20 minutes；Then, carbon printing ink is printed on Pet as working electrode with to electrode, 110 DEG C
Toast 30 minutes；Printing one layer of insulation paste again at working electrode surface again, 70 DEG C are toasted 15 minutes；Finally
Three electrodes are connected respectively conduction filamentary silver and draws circuit.
4. three-dimensional graphene electrode is surface-functionalized: weighs 2.0g chitosan and is dissolved in the acetic acid of 100ml 2.0%
In solution, room temperature stirs 3h to prepare chitosan solution with 250 revs/min；Draw 15 μ L glucose
Oxidase solution (concentration is 400IU/mL) drop coating is on GNWs, and room temperature is dried；Take 15 μ L 2.0% shells
Polysaccharide solution drop coating is to GNWs/ enzyme surface, and room temperature is dried；The glutaraldehyde drop coating taking 15 μ L 1.5% is layed onto
GNWs/ enzyme/chitosan surface, room temperature dries 1h；Electrode is immersed in the 0.1M PBS of pH 7.0,
4 DEG C overnight to remove glucoseoxidase remaining on electrode；4 DEG C of 0.1M PBS bufferings being saved in pH7.0
In liquid.
Implement row 4
The present embodiment provides the preparation method of a kind of three-dimensional grapheme glucose sensor, the technological process of the method
Figure is as it is shown in figure 1, specifically comprise the following steps that
1. use PECVD growing three-dimensional Graphene: the Copper Foil of 70 μ m-thick is lain in a horizontal plane in growth furnace
In glass tubing, successively through heating up, anneal, growing three steps.Condition is respectively as follows: intensification: 60min
It is warming up to 800 DEG C, hydrogen=30sccm；Annealing: 80min, maintains 600 DEG C, hydrogen=15sccm；Growth:
110min, maintains 650 DEG C, methane: hydrogen=6: 6sccm, radio frequency=450mw；Growth is closed after terminating and is penetrated
Frequency and methane, hydrogen arranges 12sccm, opens fan fast cooling, and the most gently taking out growth has stone
The Copper Foil of ink alkene wall.
2. three-dimensional grapheme is transferred to PET base material: prepares Fe (NO3) the 3 bronze medal etching liquid of 0.1M, then will
Copper Foil is cut to the fritter of 2.5cm*2.5cm and inserts solution surface, has the one of Graphene wall to face up with growth,
Overnight fully to etch Copper Foil under room temperature condition.Then, (the hydrophilic of pet sheet face is exposed with plasma treated
Group, convenient with the laminating of Graphene wall) 6cm*6cm PET film floating Graphene wall is picked up, by stone
Ink alkene wall is transferred in ultra-pure water remove the metal ions such as the copper of remnants, ferrum with abundant 30min*4 time.Then,
PET picks up cleaned Graphene wall and puts into drying under conditions of 100 DEG C of * 30min in baking oven, makes graphite
Alkene wall and PET secure fit.
3. use silk screen print method printed electrode: be first printed on as reference electrode on PET using Ag/AgCl ink,
80 DEG C are toasted 20 minutes；Then, carbon printing ink is printed on Pet as working electrode with to electrode, 90 DEG C
Toast 25 minutes；Printing one layer of insulation paste again at working electrode surface again, 80 DEG C are toasted 20 minutes；?
After by three electrodes respectively connect conduction filamentary silver draw circuit.
4. three-dimensional graphene electrode is surface-functionalized: weighs 3.0g chitosan and is dissolved in the acetic acid of 100ml 3.0%
In solution, room temperature stirs 3h to prepare chitosan solution with 250 revs/min；Draw 5 μ L glucose oxygen
Changing enzymatic solution (concentration is 600IU/mL) drop coating on GNWs, room temperature is dried；Take 5 μ L 3.0% shells to gather
Sugar juice drop coating is to GNWs/ enzyme surface, and room temperature is dried；The glutaraldehyde drop coating taking 5 μ L 0.5% is layed onto GNWs/
Enzyme/chitosan surface, room temperature dries 1h；Electrode is immersed in the 0.1M PBS of pH 7.0,4 DEG C
Overnight to remove glucoseoxidase remaining on electrode；4 DEG C are saved in the 0.1M PBS of pH7.0.
Implement row 5
The present embodiment provides the preparation method of a kind of three-dimensional grapheme glucose sensor, specifically comprises the following steps that
1. use PECVD growing three-dimensional Graphene: the Copper Foil of 40 μ m-thick is lain in a horizontal plane in growth furnace
In glass tubing, successively through heating up, anneal, growing three steps.Condition is respectively as follows: intensification: 40min
It is warming up to 600 DEG C, hydrogen=15sccm；Annealing: 50min, maintains 700 DEG C, hydrogen=15sccm；Growth:
80min, maintains 700 DEG C, methane: hydrogen=8: 8sccm, radio frequency=300mw；Growth closes radio frequency after terminating
And methane, hydrogen arranges 6sccm, opens fan fast cooling, and the most gently taking out growth has Graphene
The Copper Foil of wall.
2. three-dimensional grapheme is transferred to PET base material: prepare Fe (NO3) the 3 bronze medal etching liquid of 0.3M, then by copper
Paper tinsel is cut to the fritter of 1.0cm*1.0cm and inserts solution surface, has the one of Graphene wall to face up with growth, room
Overnight fully to etch Copper Foil under the conditions of temperature.Then, (hydrophilic group in pet sheet face is exposed with plasma treated
Group, convenient with the laminating of Graphene wall) 3cm*3cm PET film floating Graphene wall is picked up, by graphite
Alkene wall is transferred in ultra-pure water remove the metal ions such as the copper of remnants, ferrum with abundant 30min*5 time.Then,
PET picks up cleaned Graphene wall and puts into drying under conditions of 80 DEG C of * 60min in baking oven, makes graphite
Alkene wall and PET secure fit.
3. use silk screen print method printed electrode: be first printed on as reference electrode on PET using Ag/AgCl ink,
90 DEG C are toasted 15 minutes；Then, carbon printing ink is printed on PET as working electrode with to electrode, 100 DEG C
In the range of toast 25 minutes；Printing one layer of insulation paste again at working electrode surface again, 90 DEG C are toasted 20 points
Clock；Finally three electrodes are connected respectively conduction filamentary silver and draws circuit.
4. Plasma-Modified graphene nano wall: be divided into Normal group and Cement Composite Treated by Plasma group, adopted
Modifying graphene nano wall surface with plasma method, treatment conditions used are: oxygen flow is
30sccm, air pressure is 1mbar, 500W middle-grade process 10min；
5. three-dimensional graphene electrode is surface-functionalized: weighing 1.5g chitosan, to be dissolved in the acetic acid of 100ml 1.5% molten
In liquid, room temperature stirs 3h to prepare chitosan solution with 250 revs/min；Draw 10 μ L glucose oxygen
Changing enzymatic solution (concentration is 300IU/mL) drop coating on GNWs, room temperature is dried；Take 10, μ L 1.5% shell
Polysaccharide solution drop coating is to GNWs/ enzyme surface, and room temperature is dried；The glutaraldehyde drop coating taking 10 μ L 1.0% is layed onto
GNWs/ enzyme/chitosan surface, room temperature dries 1h；Electrode is immersed in the 0.1M PBS of pH 7.0,
4 DEG C overnight to remove glucoseoxidase remaining on electrode；4 DEG C of 0.1M PBS bufferings being saved in pH7.0
In liquid.
Claims (5)
1. a preparation method for glucose sensor, uses graphene nano wall as load skeleton, and described graphene nano wall using plasma strengthens chemical vapour deposition technique (PECVD) to be prepared, and Copper Foil successively carries out heating up, anneals, growth step；Wherein, Elevated Temperature Conditions is 30～90min to be warming up to 500～800 DEG C, hydrogen=5～30sccm；Annealing conditions is 30～90min, maintains 600～800 DEG C, hydrogen=5～15sccm；Growth conditions is 600～800 DEG C and maintains 60～120min, and methane: hydrogen is 4:8sccm, radio frequency is 200～500mw；Closing radio frequency and methane, hydrogen is set to 2～20sccm, and taking out growth after cooling has the Copper Foil of Graphene wall；After graphene nano wall is transferred to polyethylene terephthalate (PET), use after silk screen print method prints three electrodes, carry out electrode surface-functionalized glucoseoxidase loading to three-dimensional grapheme nm wall.
2. the preparation method of glucose sensor as claimed in claim 1, described surface functionalization step is: a) glucose oxidase solution of 100-800IU/ml is loaded to three-dimensional grapheme nm wall, dries；B) 0.3-3.5wt% chitosan solution is dripped it is layed onto three-dimensional grapheme/glucoseoxidase surface；C) glutaraldehyde of 0.25-2wt% is fixed to three-dimensional grapheme wall/enzyme/chitosan surface, dries, d) use PBS solution cleaning electrode and preserve.
3. the preparation method of glucose sensor as claimed in claim 2, described PBS solution concentration is 0.1M, pH=7.0, cleans and storage temperature is 2-8 DEG C.
4.
The preparation method of glucose sensor as claimed in claim 1 or 2, employing silk screen print method printed electrode step is: a) be printed on PET as reference electrode using Ag/AgCl ink, toast 5-25 minute 60-100 DEG C of scope, b) carbon printing ink is printed on PET as working electrode with to electrode, toast 15-30 minute 80-120 DEG C of scope, c) one layer of insulation paste is printed again at working electrode surface, toast in the range of 50-90 DEG C 5-20 minute, d) three electrodes are connected respectively conduction filamentary silver and draw circuit.
5. the preparation method of glucose sensor as claimed in claim 1, comprises the following steps:
(1) preparation of Graphene wall
Use PECVD, the Copper Foil that thickness is 1 μm～100 μm is lain in a horizontal plane in the glass tubing of growth furnace, successively through heating up, anneal, growing three steps；Wherein, temperature-rise period is 30～90min to be warming up to 500～800 DEG C, hydrogen=5～30sccm；Annealing process is 30～90min, maintains 600～800 DEG C, hydrogen=5～15sccm；Growth course is 60～120min, maintains 600～800 DEG C, methane: hydrogen=4:8sccm, radio frequency=200～500mw；Growth closes radio frequency and methane after terminating, and hydrogen arranges 2～20sccm, opens fan fast cooling, and the most gently taking out growth has the Copper Foil of Graphene wall；
(2) transfer of Graphene wall
A) compound concentration is the Fe (NO of 0.1M～0.5M3)3Copper etching liquid, b) Copper Foil is cut into a size of 0.2cm*0.2cm～3cm*3cm fritter not etc. and inserts solution surface, the one of Graphene wall is had to face up with growth, overnight fully to etch Copper Foil under room temperature condition, c) it is that floating Graphene wall is picked up by 0.2cm*0.2cm～3cm*3cm PET film not etc. by plasma treated size, Graphene wall is transferred to 30min*3 in ultra-pure water～5 times fully to remove remaining copper, the metal ions such as ferrum, d) pick up cleaned Graphene wall with PET and put into drying under conditions of 80～150 DEG C * 5～60min in baking oven, make Graphene wall and PET secure fit；
(3) preparation of electrode
Use silk screen print method printed electrode, a) Ag/AgCl ink is printed on PET as reference electrode, toast 5-25 minute 60-100 DEG C of scope, b) carbon printing ink is printed on PET as working electrode with to electrode, toast 15-30 minute 80-120 DEG C of scope, c) print one layer of insulation paste again at working electrode surface, toast in the range of 50-90 DEG C 5-20 minute, d) three electrodes are connected respectively conduction filamentary silver and draw circuit；
(4) sensor electrode is surface-functionalized
A) configuration concentration is the glucose oxidase solution of 100-800IU/ml, by on 2-20 μ L glucose oxidase solution loadings to three-dimensional grapheme nm wall, room temperature is dried, b) by 2-20 μ L, concentration is that 0.3-3.5% chitosan solution drips and is layed onto three-dimensional grapheme/glucoseoxidase surface, c) glutaraldehyde that concentration is 0.25-2% is fixed to three-dimensional grapheme wall/enzyme/chitosan surface, room temperature is dried a period of time, and d) remaining on cleaning electrode glucoseoxidase also preserves；During cleaning and preserving, used PBS solution concentration is 0.1M, pH=7.0, and the temperature of process is 2-8 DEG C.
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