CN105708416B - Preparation method of three-dimensional graphene lactic acid sensor - Google Patents
Preparation method of three-dimensional graphene lactic acid sensor Download PDFInfo
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- CN105708416B CN105708416B CN201610033938.2A CN201610033938A CN105708416B CN 105708416 B CN105708416 B CN 105708416B CN 201610033938 A CN201610033938 A CN 201610033938A CN 105708416 B CN105708416 B CN 105708416B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/68—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient
- A61B5/6801—Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be attached to or worn on the body surface
- A61B5/6813—Specially adapted to be attached to a specific body part
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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
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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
- G01N27/416—Systems
- G01N27/48—Systems using polarography, i.e. measuring changes in current under a slowly-varying voltage
Abstract
The invention relates to a preparation method of a three-dimensional graphene lactic acid sensor. The preparation method comprises the following steps: promoting growth of a three-dimensional graphene nano wall by virtue of a plasma-enhanced chemical vapor deposition method by taking a copper foil as a substrate; drifting a three-dimensional graphene block on the surface of an Fe(NO3)3 solution so as to etch the copper foil, cleaning the copper foil by virtue of ultra-pure water and transferring the copper foil to a polyethylene terephthalate (PET) base material; printing three electrodes on the surface of the three-dimensional graphene/PET by virtue of a screen printing technology, and immobilizing lactase oxidase on the surface of the three-dimensional graphene by virtue of a chitosan-glutaraldehyde cross-linking method, so as to conduct functionalizing treatment on the electrodes; and finally, conduct sensitivity, stability, real-time performance and specificity lactic acid detection on the electrodes with the assistance of an electrochemical workstation, so that the required three-dimensional graphene lactic acid sensor is obtained. According to the preparation method disclosed by the invention, the three-dimensional graphene lactic acid sensor is prepared in a mode of being economic and environment-friendly, simple and efficient and controllable in quality, and due to the graphene nano wall which has the advantages of being rich in three-dimensional conductive network structure, huge in specific surface area and the like, the sensitivity and the real-time performance of lactic acid detection can be effectively improved; and the sensor is applicable to the fields of medical care and health.
Description
Technical field
The present invention relates to a kind of preparation method of three-dimensional grapheme lactic acid sensor, belongs to biochemical sensor technology neck
A kind of domain, and in particular to preparation method of wearable three-dimensional grapheme lactic acid sensor.
Background technology
Lactic acid, as the product of glucose anerobic glycolysises, occurs a large amount of in the case of human body anoxia and fatigue
Accumulation, whether its content in body fluid and tissue can effectively reflect human local tissue whether anoxia, move excessive.
It can be seen that, the real-time monitoring of human body lactic acid content is particularly important.But, lactate detection instrument general on market belongs to intrusion
Formula sensor, the form pricked by pin gathers the blood of experimenter's fingerstick capillary, and collection repeatedly is not only injured greatly, and
Real-time monitoring can not be accomplished, the monitoring and therapeutic of the patient that various diseases are caused with anoxia is especially embodied in and athlete's physique is surveyed
During examination.Recently research successively reports the wearable electrochemistry for lactic acid content monitoring in human saliva and perspiration
Sensor, but its complex manufacturing process, technological requirement and high cost, precision is low, poor biocompatibility, it is impossible to timely and effectively anti-
Reflect the change of lactic acid content.Therefore, need the new lactate detection technology of research and development badly to realize that it is monitored in physiological health and signal is carried
Take the application breakthrough of aspect.
Graphene, a kind of New Two Dimensional c-based nanomaterial being made up of monolayer carbon atom, in bio electrochemistry field
Research it is of increasing concern, its excellent electrocatalysis characteristic becomes and prepares the preferable electrode of enzyme electrochemica biological sensor
One of material, its special molecular structure allows it to be effectively facilitated electric transmission, lifts the response signal of biosensor
And sensitivity.Additionally, its good biocompatibility can keep sensor to load the biological activity of enzyme, improve sensor
Stability.Recently, graphene nano wall, a kind of Graphene by graphene nanometer sheet vertical distribution on base material is three-dimensional to be tied
Structure, the morphosiss with more horn of plenty and huge specific surface area compared with two-dimensional graphene, while imparting preferably drawing
Stretching property, has started to gradually be applied to the research and development of sensor, such as to gas, the monitoring of temperature.
Application publication number discloses a kind of lactic biological sensor of graphene modified for CN105116028A patent documents
And preparation method thereof, the lactic biological sensor is a kind of lactic acid sensor of miniaturization, the lactic acid in measurable human serum
Content, but the material still belongs to the Wearable lactic acid sensor of the carbon granule/carbon fiber for base material of two dimension, its conductivity
Property, biocompatibility still have limitation, and material tensile property itself is not strong.Simultaneously the patent uses oxidation or carboxyl
The Graphene of change, material itself causes its electric transmission speed not high due to its fault of construction, affects the efficiency of sensor.
The content of the invention
For the not enough and Improvement requirement in background technology, it is an object of the invention to provide a kind of three-dimensional grapheme lactic acid
The preparation method of sensor, based on a kind of special graphene nano wall construction material as Lactate Oxidase load skeleton,
Three-dimensional grapheme has higher electrical conductivity, the conductive network structure of more horn of plenty and material bio-compatibility is good.By right
The growth conditionss research of key sensor base material-three-dimensional grapheme wall and technological improvement, the preparation and lactalase work(to sensor
Technical optimization can be changed, new three-dimensional grapheme lactic acid sensor, and the sensor can be obtained for the electrochemistry sound of lactic acid
Should be sensitive, and excellent repeatability is embodied, while tensility is strong, suitable wearing, this sensor can be used on Noninvasive
Wearable electronic skin, can carry out real-time, accurate detection to human body lactic acid；This three-dimensional grapheme is with low cost, product
Product simple production process, is suitable to large-scale industrialization batch production, is particularly suited for the fields such as medical monitoring, health control
Using.A kind of preparation method of three-dimensional grapheme lactic acid sensor involved in the present invention, the method step is as follows：
(1) preparation of Graphene wall
Using PECVD, the Copper Foil that thickness is 1 μm～100 μm is lain in a horizontal plane in the glass tubing of growth furnace, Jing after elder generation
Cross intensification, annealing, three steps of growth；Wherein, temperature-rise period is warming up to 500～800 DEG C for 30～90min, and hydrogen=5～
30sccm；Annealing process is 30～90min, maintains 600～800 DEG C, hydrogen=5～15sccm；Growth course be 60～
120min, maintains 600～800 DEG C, methane:Hydrogen=4:8sccm, radio frequency=200～500mw；Growth closes radio frequency after terminating
And methane, hydrogen setting to 2～20sccm, opening fan fast cooling, finally gently taking out growth has the copper of Graphene wall
Paper tinsel；
(2) transfer of Graphene wall
A) compound concentration is the Fe (NO of 0.1M～0.5M3)3Copper etching liquid, b) is cut into size for 0.2cm* by Copper Foil
The fritter that 0.2cm～3cm*3cm is not waited inserts solution surface, is faced up with growing have Graphene wall one, and room temperature condition was descended
Night, fully to etch Copper Foil, is c) that the PET film that 0.2cm*0.2cm～3cm*3cm is not waited will float with plasma treated size
Floating Graphene wall is picked up, and Graphene wall is transferred to into 30min*3～5 time in ultra-pure water and removes remaining copper, ferrum etc. with abundant
Metal ion, the Graphene wall that cleaned d) is picked up with PET and is put in baking oven under conditions of 80～150 DEG C of * 5～60min
Drying, makes Graphene wall and PET secure fits；
(3) preparation of electrode
Using silk screen print method printed electrode, a) Ag/AgCl ink is printed on PET as reference electrode, in 60-
100 DEG C of scopes toast 5-25 minutes, b) are printed on carbon printing ink on PET as working electrode and to electrode, in 80-120 DEG C of model
Baking 15-30 minutes are enclosed, c) one layer of insulation paste is printed again in working electrode surface, 5-20 point is toasted in the range of 50-90 DEG C
Three electrodes d) are connected respectively conductive filamentary silver and draw circuit by clock；
(4) sensor electrode is surface-functionalized
A) Lactate Oxidase is loaded on three-dimensional grapheme nm wall, room temperature is dried, b) configures 2-20 μ L, concentration is
The chitosan solution of 100-800IU/ml, c) by 2-20 μ L, concentration is that 0.3-3.5% chitosan solutions drop is layed onto three-dimensional graphite
Alkene/Lactate Oxidase surface, d) is fixed to three-dimensional grapheme wall/enzyme/shitosan surface by the glutaraldehyde that concentration is 0.25-2%,
Room temperature dries a period of time, e) Lactate Oxidase remaining on cleaning electrode and preserves；It is used during cleaning and preservation
To PBS solution concentration be 0.1M, pH=7.0, the temperature of process is 2-8 DEG C；
(5) lactate detection of sensor electrode
A) the making lactic acid solution in PBS, b) tests transducer sensitivity and tests sensor stability, c) surveys
Examination sensor specificity；Wherein, the lactic acid solution concentration range that the PBS that concentration is 0.1M, pH value is 7.0 is prepared is in 1-
100mM；In oxidation-reduction curve test process, potential is set as into -1.0V-+1.0V (VS.Ag/AgCl), by sensor
Successively it is immersed in above-mentioned solution, start recording electric current after 2min, the record time is 60s；In time current curve test process
In, potential is set as into -0.35V (VS.Ag/AgCl), sensor is immersed in above-mentioned solution and records the time for 60s；In spirit
In sensitivity test process, electrode is immersed in successively in the lactic acid solution of 0.1-400mM after 2min, connects corresponding three electricity
Pole, opens electrochemical workstation (CHI760), and potential is set as into -0.35V (VS.Ag/AgCl), is started using cyclic voltammetry
Record current, the record time is 400s.
In general, possess following compared with prior art, mainly according to the contemplated above technical scheme of the present invention
Technological merit：
1. with three-dimensional grapheme nm wall as base material, three electrodes are printed using silk screen print method, then the shell by optimizing
Lactate Oxidase is fixed to nm wall surface and electrode is obtained by polysaccharide-glutaraldehyde cross-linking absorption method.Electrode test shows：With this
Electrode prepared by method, with good susceptiveness, stability and real-time, while there is preferable lactic acid identification specificity；
2. compared with two-dimensional graphene, the graphene nano wall that the invention is used has more abundant three dimensional structure, more
For huge specific surface area and more superior electron transfer performance, more Lactate Oxidase, and structure have not only been loaded
A more accurate conductive network is built, so as to effectively improve the sensitivity to lactate detection；
3. the method prepares lactic acid sensor, and accurate real-time, stable performance, biocompatibility are strong, economic and environment-friendly, and technique
Simply, it is with low cost, without the need for precise control technological parameter, be suitable to large-scale industrialization batch production, be particularly suited for medical treatment
The application in the fields such as monitoring, health control.
Description of the drawings
Fig. 1 is a kind of preparation technology flow chart of three-dimensional grapheme lactic acid sensor involved in the present invention；
Fig. 2 is three-dimensional grapheme nm wall high-resolution-ration transmission electric-lens photo (A) obtained by embodiment 1 and different times magnifications
Number stereoscan photograph (B-E)；
Fig. 3 is the Raman spectrum characteristic peak of the three-dimensional grapheme nm wall obtained by embodiment 1；
Fig. 4 is the oxidoreduction curve of the three-dimensional grapheme lactic acid sensor product obtained by embodiment 1, PBS be without
The phosphate buffer of lactic acid；
Fig. 5 is the time current curve of the three-dimensional grapheme lactic acid sensor product obtained by embodiment 1, PBS be without
The phosphate buffer of lactic acid；
Fig. 6 is the electricity of three-dimensional grapheme lactic acid sensor product obtained by embodiment 1 in variable concentrations lactic acid solution
Stream change curve.Initial lactic acid concn is 0.5mM, then sensor is respectively implanted into a solution (0.6mM), b solution (0.7mM),
C solution (0.8mM), d solution (0.1mM), e solution (0.8mM) and f solution (0mM), the current value size for being recorded is with solution
Lactic acid concn size is proportionate.
Specific embodiment
Below in conjunction with the accompanying drawings the present invention will be described with row are implemented.
Implement row 1
The present embodiment provides a kind of preparation method of three-dimensional grapheme lactic acid sensor, and the process chart of the method is as schemed
Shown in 1, comprise the following steps that：
1. PECVD is adopted, the Copper Foil of 20 μ m-thicks is lain in a horizontal plane in the glass tubing of growth furnace, successively process intensification,
Annealing, three steps of growth.Condition is respectively：Heat up：30min is warming up to 500 DEG C, hydrogen=5, and 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；Radio frequency and methane are closed in growth after terminating, hydrogen is arranged to 2sccm, opens fan fast cooling,
Last gently taking-up grows the Copper Foil for having Graphene wall, obtained three-dimensional grapheme nm wall high-resolution-ration transmission electric-lens photo
And different amplification stereoscan photograph (B-E) is as shown in Fig. 2 the Raman light of obtained three-dimensional grapheme nm wall (A)
Spectrum signature peak is as shown in Figure 3.
2. Fe (the NO of 0.1M are prepared3)3Copper etching liquid, then inserts solution by the fritter that Copper Foil is cut to 0.5cm*0.5cm
Surface, to grow facing up of having Graphene wall, overnight fully etching Copper Foil under room temperature condition.Then, Jing plasmas are used
Process the Graphene wall that the 1cm*1cm PET films of (hydrophilic group in exposure pet sheet face, convenient to fit with Graphene wall) will be floated
Pick up, Graphene wall is transferred in ultra-pure water 30min*3 time fully to remove the metal ions such as remaining copper, ferrum.Then,
PET picks up the Graphene wall that cleaned and is put in baking oven and toasts 5min under conditions of 80 DEG C, makes Graphene wall firm with PET
Laminating.
3. silk screen print method printed electrode is adopted, first Ag/AgCl ink is printed on as reference electrode on PET, 70 DEG C
Baking 5 minutes；Then, carbon printing ink is printed on as working electrode and to electrode on PET, 80 DEG C are toasted 15 minutes；Again in work
Make electrode surface and print one layer of insulation paste again, 70 DEG C are toasted 10 minutes；Finally three electrodes are connected respectively into conductive filamentary silver to draw
Circuit.
4. weigh 0.5g shitosans to be dissolved in the acetic acid solution of 100ml 0.5%, room temperature with 250 revs/min stirring 3h with
Prepare chitosan solution；To on GNWs, room temperature is dried to draw 3 μ L Lox solution (concentration is 100IU/mL) drop coatings；Take 3 μ L
0.5% chitosan solution drop coating to GNWs/ enzymes surface, dry by room temperature；Take 3 μ L0.25% glutaraldehyde drop coating be layed onto GNWs/ enzymes/
Shitosan surface, room temperature dries 1h；Electrode is immersed in the 0.1M PBSs of pH 7.0,4 DEG C overnight removing electrode
Upper remaining Lactate Oxidase；4 DEG C are stored in the 0.1M PBSs of pH7.0.
5. in concentration be 0.1M, the PBS that pH value is 7.0 in compound concentration for 1mM lactic acid solution；Oxidation-
Also in virgin curve test process, potential is set as into -1.0V-+1.0V (VS.Ag/AgCl), sensor is successively immersed in above-mentioned
In solution, start recording electric current after 2min, the record time is 60s；In time current curve test process, potential is set
For -0.35V (VS.Ag/AgCl), sensor is immersed in above-mentioned solution and records the time for 60s；In product sensitivity test
Cheng Zhong, compound concentration is the breast of 0mM, 0.1mM, 0.5mM, 0.6mM, 0.7mM, 0.8mM in PBS (0.1M, pH7.0) buffer
Acid solution, by potential -0.35V (VS.Ag/AgCl) is set as, sensor is successively immersed in above-mentioned solution, and the record time is
400s.Fig. 4 is the oxidoreduction curve of the three-dimensional grapheme lactic acid sensor product obtained by embodiment 1, and PBS is without lactic acid
Phosphate buffer；The time current curve of obtained three-dimensional grapheme lactic acid sensor product is as shown in figure 5, made
Curent change curve of the three-dimensional grapheme lactic acid sensor product for obtaining in variable concentrations lactic acid solution is as shown in Figure 6.
Implement row 2
The present embodiment provides a kind of preparation method of three-dimensional grapheme lactic acid sensor, and the process chart of the method is as schemed
Shown in 1, comprise the following steps that：
1. PECVD is adopted, the Copper Foil of 30 μ m-thicks is lain in a horizontal plane in the glass tubing of growth furnace, successively process intensification,
Annealing, three steps of growth.Condition is respectively：Heat up：35min is warming up to 550 DEG C, hydrogen=5, and 10,15,20,25,
30sccm；Annealing：40min, maintains 650 DEG C, hydrogen=10sccm；Growth：70min, maintains 650 DEG C, methane:Hydrogen=6:
8sccm, radio frequency=250mw；Radio frequency and methane are closed in growth after terminating, hydrogen is arranged to 4sccm, opens fan fast cooling,
Last gently taking-up grows the Copper Foil for having Graphene wall.
2. Fe (the NO of 0.2M are prepared3)3Copper etching liquid, then inserts solution by the fritter that Copper Foil is cut to 0.8cm*0.8cm
Surface, to grow facing up of having Graphene wall, overnight fully etching Copper Foil under room temperature condition.Then, Jing plasmas are used
Process the Graphene wall that the 2cm*2cm PET films of (hydrophilic group in exposure pet sheet face, convenient to fit with Graphene wall) will be floated
Pick up, Graphene wall is transferred in ultra-pure water 30min*4 time fully to remove the metal ions such as remaining copper, ferrum.Then,
PET picks up the Graphene wall that cleaned and is put in baking oven and dries under conditions of 80 DEG C of * 20min, makes Graphene wall firm with PET
Gu laminating.
3. silk screen print method printed electrode (as shown in Figure 1) is adopted.First Ag/AgCl ink is printed on PET as ginseng
Than electrode, 80 DEG C are toasted 10 minutes；Then, carbon printing ink is printed on as working electrode and to electrode on Pet, 90 DEG C of bakings 20
Minute；Again one layer of insulation paste is printed again in working electrode surface, 80 DEG C are toasted 15 minutes；Finally three electrodes are connected respectively and is led
Electric filamentary silver draws circuit.
4. weigh 1g shitosans to be dissolved in the acetic acid solution of 100ml 1.0%, room temperature stirs 3h to match somebody with somebody with 250 revs/min
Chitosan solution processed；To on GNWs, room temperature is dried to draw 5 μ L Lox solution (concentration is 200IU/mL) drop coatings；Take 5 μ L 1.0%
Chitosan solution drop coating to GNWs/ enzymes surface, dry by room temperature；Take 5 μ L 0.5% glutaraldehyde drop coating be layed onto GNWs/ enzymes/shell gather
Sugared surface, room temperature dries 1h；Electrode is immersed in the 0.1M PBSs of pH 7.0,4 DEG C overnight residual on electrode to remove
Remaining Lactate Oxidase；4 DEG C are stored in the 0.1M PBSs of pH7.0.
5. in concentration be 0.1M, the PBS that pH value is 7.0 in compound concentration for 5mM lactic acid solution；Oxidation-
Also in virgin curve test process, potential is set as into -1.0V-+1.0V (VS.Ag/AgCl), sensor is successively immersed in above-mentioned
In solution, start recording electric current after 2min, the record time is 60s；In time current curve test process, potential is set
For -0.35V (VS.Ag/AgCl), sensor is immersed in above-mentioned solution and records the time for 60s；In product sensitivity test
Cheng Zhong, compound concentration is the lactic acid of 0mM, 1mM, 2.0mM, 4.0mM, 6.0mM, 8.0mM in PBS (0.1M, pH7.0) buffer
Solution, by potential -0.35V (VS.Ag/AgCl) is set as, sensor is successively immersed in above-mentioned solution, and the record time is
400s。
Implement row 3
The present embodiment provides a kind of preparation method of three-dimensional grapheme lactic acid sensor, and the process chart of the method is as schemed
Shown in 1, comprise the following steps that：
1. PECVD is adopted, the Copper Foil of 40 μ m-thicks is lain in a horizontal plane in the glass tubing of growth furnace, successively process intensification,
Annealing, three steps of growth.Condition is respectively：Heat up：40min is warming up to 600 DEG C, hydrogen=15sccm；Annealing：50min, dimension
700 DEG C are held, hydrogen=15sccm；Growth：80min, maintains 700 DEG C, methane:Hydrogen=8：8sccm, radio frequency=300mw；Growth
Radio frequency and methane are closed after end, hydrogen is arranged to 6sccm, opens fan fast cooling, finally gently taking out growth has stone
The Copper Foil of black alkene wall.
2. Fe (the NO of 0.3M are prepared3)3Copper etching liquid, then inserts solution by the fritter that Copper Foil is cut to 1.0cm*1.0cm
Surface, to grow facing up of having Graphene wall, overnight fully etching Copper Foil under room temperature condition.Then, Jing plasmas are used
Process the Graphene wall that the 3cm*3cm PET films of (hydrophilic group in exposure pet sheet face, convenient to fit with Graphene wall) will be floated
Pick up, Graphene wall is transferred in ultra-pure water 30min*5 time fully to remove the metal ions such as remaining copper, ferrum.Then,
PET picks up the Graphene wall that cleaned and is put in baking oven and dries under conditions of 80 DEG C of * 60min, makes Graphene wall firm with PET
Gu laminating.
3. silk screen print method printed electrode (as shown in Figure 1) is adopted.First Ag/AgCl ink is printed on PET as ginseng
Than electrode, 90 DEG C are toasted 15 minutes；Then, carbon printing ink is printed on as working electrode and to electrode on PET, 100 DEG C of scopes
Interior baking 25 minutes；Again one layer of insulation paste is printed again in working electrode surface, 90 DEG C are toasted 20 minutes；Finally by three electrodes point
Do not connect conductive filamentary silver and draw circuit.
4. weigh 1.5g shitosans to be dissolved in the acetic acid solution of 100ml 1.5%, room temperature with 250 revs/min stirring 3h with
Prepare chitosan solution；To on GNWs, room temperature is dried to draw 10 μ L Lox solution (concentration is 300IU/mL) drop coatings；Take 10, μ L
1.5% chitosan solution drop coating to GNWs/ enzymes surface, dry by room temperature；The glutaraldehyde drop coating for taking 10 μ L 1.0% is layed onto GNWs/
Enzyme/shitosan surface, room temperature dries 1h；Electrode is immersed in the 0.1M PBSs of pH 7.0,4 DEG C overnight removing electricity
Extremely go up the Lactate Oxidase of remnants；4 DEG C are stored in the 0.1M PBSs of pH7.0.
5. in concentration be 0.1M, the PBS that pH value is 7.0 in compound concentration for 10mM lactic acid solution；Oxidation-
Also in virgin curve test process, potential is set as into -1.0V-+1.0V (VS.Ag/AgCl), sensor is successively immersed in above-mentioned
In solution, start recording electric current after 2min, the record time is 60s；In time current curve test process, potential is set
For -0.35V (VS.Ag/AgCl), sensor is immersed in above-mentioned solution and records the time for 60s；In product sensitivity test
Cheng Zhong, compound concentration is molten for the lactic acid of 0mM, 0.1mM, 1mM, 10mM, 20mM, 40mM in PBS (0.1M, pH7.0) buffer
Liquid, by potential -0.35V (VS.Ag/AgCl) is set as, sensor is successively immersed in above-mentioned solution, and the record time is
400s。
Implement row 4
The present embodiment provides a kind of preparation method of three-dimensional grapheme lactic acid sensor, and the process chart of the method is as schemed
Shown in 1, comprise the following steps that：
1. PECVD is adopted, the Copper Foil of 50 μ m-thicks is lain in a horizontal plane in the glass tubing of growth furnace, successively process intensification,
Annealing, three steps of growth.Condition is respectively：Heat up：45min is warming up to 650 DEG C, hydrogen=20sccm；Annealing：60min, dimension
750 DEG C are held, hydrogen=5sccm；Growth：90min, maintains 750 DEG C, methane:Hydrogen=10：8sccm, radio frequency=350mw；Growth
Radio frequency and methane are closed after end, hydrogen is arranged to 8sccm, opens fan fast cooling, finally gently taking out growth has stone
The Copper Foil of black alkene wall.
2. Fe (the NO of 0.4.M are prepared3)3Copper etching liquid, then inserts the fritter that Copper Foil is cut to 1.5cm*1.5cm molten
Liquid surface, to grow facing up of having Graphene wall, overnight fully etching Copper Foil under room temperature condition.Then, with Jing etc. from
The Graphene that the 4cm*4cm PET films of subprocessing (hydrophilic group in exposure pet sheet face, convenient to fit with Graphene wall) will be floated
Wall is picked up, and Graphene wall is transferred in ultra-pure water 30min*3 time fully to remove the metal ions such as remaining copper, ferrum.Then,
PET picks up the Graphene wall that cleaned and is put in baking oven and dries under conditions of 100*10min, makes Graphene wall firm with PET
Gu laminating.
3. silk screen print method printed electrode (as shown in Figure 1) is adopted.First Ag/AgCl ink is printed on PET as ginseng
Than electrode, 100 DEG C are toasted 20 minutes；Then, carbon printing ink is printed on as working electrode and to electrode on Pet, 110 DEG C of bakings
30 minutes；Again one layer of insulation paste is printed again in working electrode surface, 70 DEG C are toasted 15 minutes；Finally three electrodes are connected respectively
Conductive filamentary silver draws circuit.
4. weigh 2.0g shitosans to be dissolved in the acetic acid solution of 100ml 2.0%, room temperature with 250 revs/min stirring 3h with
Prepare chitosan solution；To on GNWs, room temperature is dried to draw 15 μ L Lox solution (concentration is 400IU/mL) drop coatings；Take 15 μ L
2.0% chitosan solution drop coating to GNWs/ enzymes surface, dry by room temperature；The glutaraldehyde drop coating for taking 15 μ L 1.5% is layed onto GNWs/
Enzyme/shitosan surface, room temperature dries 1h；Electrode is immersed in the 0.1M PBSs of pH 7.0,4 DEG C overnight removing electricity
Extremely go up the Lactate Oxidase of remnants；4 DEG C are stored in the 0.1M PBSs of pH7.0.
5. in concentration be 0.1M, the PBS that pH value is 7.0 in compound concentration for 20mM lactic acid solution；Oxidation-
Also in virgin curve test process, potential is set as into -1.0V-+1.0V (VS.Ag/AgCl), sensor is successively immersed in above-mentioned
In solution, start recording electric current after 2min, the record time is 60s；In time current curve test process, potential is set
For -0.35V (VS.Ag/AgCl), sensor is immersed in above-mentioned solution and records the time for 60s；In product sensitivity test
Cheng Zhong, compound concentration is molten for the lactic acid of 0mM, 1mM, 10mM, 20mM, 40mM, 60mM in PBS (0.1M, pH7.0) buffer
Liquid, by potential -0.35V (VS.Ag/AgCl) is set as, sensor is successively immersed in above-mentioned solution, and the record time is
400s。
Implement row 5
The present embodiment provides a kind of preparation method of three-dimensional grapheme lactic acid sensor, and the process chart of the method is as schemed
Shown in 1, comprise the following steps that：
1. PECVD is adopted, the Copper Foil of 60 μ m-thicks is lain in a horizontal plane in the glass tubing of growth furnace, successively process intensification,
Annealing, three steps of growth.Condition is respectively：Heat up：50min is warming up to 700 DEG C, hydrogen=25sccm；Annealing：70min, dimension
750 DEG C are held, hydrogen=10sccm；Growth：100min, maintains 800 DEG C, methane:Hydrogen=9：6sccm, radio frequency=400mw；It is raw
Length closes radio frequency and methane after terminating, hydrogen is arranged to 10sccm, opens fan fast cooling, and finally gently taking out growth has
The Copper Foil of Graphene wall.
2. Fe (the NO of 0.5M are prepared3)3Copper etching liquid, then inserts solution by the fritter that Copper Foil is cut to 2.0cm*2.0cm
Surface, to grow facing up of having Graphene wall, overnight fully etching Copper Foil under room temperature condition.Then, Jing plasmas are used
Process the Graphene wall that the 4cm*5cm PET films of (hydrophilic group in exposure pet sheet face, convenient to fit with Graphene wall) will be floated
Pick up, Graphene wall is transferred in ultra-pure water 30min*4 time fully to remove the metal ions such as remaining copper, ferrum.Then,
PET picks up the Graphene wall that cleaned and is put in baking oven and dries under conditions of 100 DEG C of * 15min, makes Graphene wall and PET
Secure fit.
3. silk screen print method printed electrode (as shown in Figure 1) is adopted.First Ag/AgCl ink is printed on PET as ginseng
Than electrode, 70 DEG C are toasted 10 minutes；Then, carbon printing ink is printed on as working electrode and to electrode on PET, 120 DEG C of bakings
15 minutes；Again one layer of insulation paste is printed again in working electrode surface, 80 DEG C are toasted 10 minutes；Finally three electrodes are connected respectively
Conductive filamentary silver draws circuit.
4. weigh 2.5g shitosans to be dissolved in the acetic acid solution of 100ml 2.5%, room temperature with 250 revs/min stirring 3h with
Prepare chitosan solution；To on GNWs, room temperature is dried to draw 20 μ L Lox solution (concentration is 500IU/mL) drop coatings；Take 20 μ L
2.5% chitosan solution drop coating to GNWs/ enzymes surface, dry by room temperature；The glutaraldehyde drop coating for taking 20 μ L 2.0% is layed onto GNWs/
Enzyme/shitosan surface, room temperature dries 1h；Electrode is immersed in the 0.1M PBSs of pH 7.0,4 DEG C overnight removing electricity
Extremely go up the Lactate Oxidase of remnants；4 DEG C are stored in the 0.1M PBSs of pH7.0.
5. in concentration be 0.1M, the PBS that pH value is 7.0 in compound concentration for 30mM lactic acid solution；Oxidation-
Also in virgin curve test process, potential is set as into -1.0V-+1.0V (VS.Ag/AgCl), sensor is successively immersed in above-mentioned
In solution, start recording electric current after 2min, the record time is 60s；In time current curve test process, potential is set
For -0.35V (VS.Ag/AgCl), sensor is immersed in above-mentioned solution and records the time for 60s；In product sensitivity test
Cheng Zhong, compound concentration is the breast of 0mM, 0.5mM, 2.0mM, 6.0mM, 7.0mM, 8.0mM in PBS (0.1M, pH7.0) buffer
Acid solution, by potential -0.35V (VS.Ag/AgCl) is set as, sensor is successively immersed in above-mentioned solution, and the record time is
400s。
Implement row 6
The present embodiment provides a kind of preparation method of three-dimensional grapheme lactic acid sensor, and the process chart of the method is as schemed
Shown in 1, comprise the following steps that：
1. PECVD is adopted, the Copper Foil of 70 μ m-thicks is lain in a horizontal plane in the glass tubing of growth furnace, successively process intensification,
Annealing, three steps of growth.Condition is respectively：Heat up：60min is warming up to 800 DEG C, hydrogen=30sccm；Annealing：80min, dimension
600 DEG C are held, hydrogen=15sccm；Growth：110min, maintains 650 DEG C, methane:Hydrogen=6：6sccm, radio frequency=450mw；It is raw
Length closes radio frequency and methane after terminating, hydrogen is arranged to 12sccm, opens fan fast cooling, and finally gently taking out growth has
The Copper Foil of Graphene wall.
2. Fe (the NO of 0.1M are prepared3)3Copper etching liquid, then inserts solution by the fritter that Copper Foil is cut to 2.5cm*2.5cm
Surface, to grow facing up of having Graphene wall, overnight fully etching Copper Foil under room temperature condition.Then, Jing plasmas are used
Process the Graphene wall that the 6cm*6cm PET films of (hydrophilic group in exposure pet sheet face, convenient to fit with Graphene wall) will be floated
Pick up, Graphene wall is transferred in ultra-pure water 30min*4 time fully to remove the metal ions such as remaining copper, ferrum.Then,
PET picks up the Graphene wall that cleaned and is put in baking oven and dries under conditions of 100 DEG C of * 30min, makes Graphene wall and PET
Secure fit.
3. silk screen print method printed electrode (as shown in Figure 1) is adopted.First Ag/AgCl ink is printed on PET as ginseng
Than electrode, 80 DEG C are toasted 20 minutes；Then, carbon printing ink is printed on as working electrode and to electrode on Pet, 90 DEG C of bakings 25
Minute；Again one layer of insulation paste is printed again in working electrode surface, 80 DEG C are toasted 20 minutes；Finally three electrodes are connected respectively and is led
Electric filamentary silver draws circuit.
4. weigh 3.0g shitosans to be dissolved in the acetic acid solution of 100ml 3.0%, room temperature with 250 revs/min stirring 3h with
Prepare chitosan solution；To on GNWs, room temperature is dried to draw 5 μ L Lox solution (concentration is 600IU/mL) drop coatings；Take 5 μ L
3.0% chitosan solution drop coating to GNWs/ enzymes surface, dry by room temperature；Take 5 μ L 0.5% glutaraldehyde drop coating be layed onto GNWs/ enzymes/
Shitosan surface, room temperature dries 1h；Electrode is immersed in the 0.1M PBSs of pH 7.0,4 DEG C overnight removing electrode
Upper remaining Lactate Oxidase；4 DEG C are stored in the 0.1M PBSs of pH7.0.
5. in concentration be 0.1M, the PBS that pH value is 7.0 in compound concentration for 40mM lactic acid solution；Oxidation-
Also in virgin curve test process, potential is set as into -1.0V-+1.0V (VS.Ag/AgCl), sensor is successively immersed in above-mentioned
In solution, start recording electric current after 2min, the record time is 60s；In time current curve test process, potential is set
For -0.35V (VS.Ag/AgCl), sensor is immersed in above-mentioned solution and records the time for 60s；In product sensitivity test
Cheng Zhong, compound concentration is the lactic acid of 0mM, 0.5mM, 10mM, 60mM, 80mM, 100mM in PBS (0.1M, pH7.0) buffer
Solution, by potential -0.35V (VS.Ag/AgCl) is set as, sensor is successively immersed in above-mentioned solution, and the record time is
400s。
Implement row 7
The present embodiment provides a kind of preparation method of three-dimensional grapheme lactic acid sensor, and the process chart of the method is as schemed
Shown in 1, comprise the following steps that：
1. PECVD is adopted, the Copper Foil of 80 μ m-thicks is lain in a horizontal plane in the glass tubing of growth furnace, successively process intensification,
Annealing, three steps of growth.Condition is respectively：Heat up：70min is warming up to 500 DEG C, hydrogen=10sccm；Annealing：90min, dimension
650 DEG C are held, hydrogen=10sccm；Growth：120min, maintains 700 DEG C, methane:Hydrogen=8：4sccm, radio frequency=300mw；It is raw
Length closes radio frequency and methane after terminating, hydrogen is arranged to 14sccm, opens fan fast cooling, and finally gently taking out growth has
The Copper Foil of Graphene wall.
2. Fe (the NO of 0.3M are prepared3)3Copper etching liquid, then inserts solution by the fritter that Copper Foil is cut to 3.0cm*3.0cm
Surface, to grow facing up of having Graphene wall, overnight fully etching Copper Foil under room temperature condition.Then, Jing plasmas are used
Process the Graphene wall that the 6cm*6cm PET films of (hydrophilic group in exposure pet sheet face, convenient to fit with Graphene wall) will be floated
Pick up, Graphene wall is transferred in ultra-pure water 30min*5 time fully to remove the metal ions such as remaining copper, ferrum.Then,
PET picks up the Graphene wall that cleaned and is put in baking oven and dries under conditions of 120*5min, makes Graphene wall firm with PET
Laminating.
3. silk screen print method printed electrode (as shown in Figure 1) is adopted.First Ag/AgCl ink is printed on PET as ginseng
Than electrode, 100 DEG C are toasted 25 minutes；Then, carbon printing ink is printed on as working electrode and to electrode on Pet, 110 DEG C of bakings
20 minutes；Again one layer of insulation paste is printed again in working electrode surface, 90 DEG C are toasted 10 minutes；Finally three electrodes are connected respectively
Conductive filamentary silver draws circuit.
4. weigh 1.2g shitosans to be dissolved in the acetic acid solution of 100ml 1.5%, room temperature with 250 revs/min stirring 3h with
Prepare chitosan solution；To on GNWs, room temperature is dried to draw 15 μ L Lox solution (concentration is 700IU/mL) drop coatings；Take 15 μ L
1.2% chitosan solution drop coating to GNWs/ enzymes surface, dry by room temperature；Take 5 μ L 1.5% glutaraldehyde drop coating be layed onto GNWs/ enzymes/
Shitosan surface, room temperature dries 1h；Electrode is immersed in the 0.1M PBSs of pH 7.0,4 DEG C overnight removing electrode
Upper remaining Lactate Oxidase；4 DEG C are stored in the 0.1M PBSs of pH7.0.
5. in concentration be 0.1M, the PBS that pH value is 7.0 in compound concentration for 60mM lactic acid solution；Oxidation-
Also in virgin curve test process, potential is set as into -1.0V-+1.0V (VS.Ag/AgCl), sensor is successively immersed in above-mentioned
In solution, start recording electric current after 2min, the record time is 60s；In time current curve test process, potential is set
For -0.35V (VS.Ag/AgCl), sensor is immersed in above-mentioned solution and records the time for 60s；In product sensitivity test
Cheng Zhong, compound concentration is the lactic acid of 0mM, 0.5mM, 20mM, 80mM, 160mM, 240mM in PBS (0.1M, pH7.0) buffer
Solution, by potential -0.35V (VS.Ag/AgCl) is set as, sensor is successively immersed in above-mentioned solution, and the record time is
400s。
Implement row 8
The present embodiment provides a kind of preparation method of three-dimensional grapheme lactic acid sensor, and the process chart of the method is as schemed
Shown in 1, comprise the following steps that：
1. PECVD is adopted, the Copper Foil of 100 μ m-thicks is lain in a horizontal plane in the glass tubing of growth furnace, successively process intensification,
Annealing, three steps of growth.Condition is respectively：Heat up：90min is warming up to 750 DEG C, hydrogen=15sccm；Annealing：60min, dimension
750 DEG C are held, hydrogen=10sccm；Growth：90min, maintains 700 DEG C, methane:Hydrogen=12：8sccm, radio frequency=350mw；It is raw
Length closes radio frequency and methane after terminating, hydrogen is arranged to 20sccm, opens fan fast cooling, and finally gently taking out growth has
The Copper Foil of Graphene wall.
2. Fe (the NO of 0.4M are prepared3)3Copper etching liquid, then inserts solution by the fritter that Copper Foil is cut to 1.0cm*2.0cm
Surface, to grow facing up of having Graphene wall, overnight fully etching Copper Foil under room temperature condition.Then, Jing plasmas are used
Process the Graphene wall that the 3cm*4cm PET films of (hydrophilic group in exposure pet sheet face, convenient to fit with Graphene wall) will be floated
Pick up, Graphene wall is transferred in ultra-pure water 30min*5 time fully to remove the metal ions such as remaining copper, ferrum.Then,
PET picks up the Graphene wall that cleaned and is put in baking oven and dries under conditions of 120*20min, makes Graphene wall firm with PET
Gu laminating.
3. silk screen print method printed electrode (as shown in Figure 1) is adopted.First Ag/AgCl ink is printed on PET as ginseng
Than electrode, 80 DEG C are toasted 25 minutes；Then, carbon printing ink is printed on as working electrode and to electrode on PET, 80 DEG C of bakings 30
Minute；Again one layer of insulation paste is printed again in working electrode surface, 70 DEG C are toasted 20 minutes；Finally three electrodes are connected respectively and is led
Electric filamentary silver draws circuit.
4. weigh 2.2g shitosans to be dissolved in the acetic acid solution of 100ml 1.5%, room temperature with 250 revs/min stirring 3h with
Prepare chitosan solution；To on GNWs, room temperature is dried to draw 3 μ L Lox solution (concentration is 400IU/mL) drop coatings；Take 10 μ L
2.2% chitosan solution drop coating to GNWs/ enzymes surface, dry by room temperature；The glutaraldehyde drop coating for taking 15 μ L 1.5% is layed onto GNWs/
Enzyme/shitosan surface, room temperature dries 1h；Electrode is immersed in the 0.1M PBSs of pH 7.0,4 DEG C overnight removing electricity
Extremely go up the Lactate Oxidase of remnants；4 DEG C are stored in the 0.1M PBSs of pH7.0.
5. in concentration be 0.1M, the PBS that pH value is 7.0 in compound concentration for 80mM lactic acid solution；Oxidation-
Also in virgin curve test process, potential is set as into -1.0V-+1.0V (VS.Ag/AgCl), sensor is successively immersed in above-mentioned
In solution, start recording electric current after 2min, the record time is 60s；In time current curve test process, potential is set
For -0.35V (VS.Ag/AgCl), sensor is immersed in above-mentioned solution and records the time for 60s；In product sensitivity test
Cheng Zhong, compound concentration is the lactic acid of 0mM, 1mM, 100mM, 150mM, 200mM, 250mM in PBS (0.1M, pH7.0) buffer
Solution, by potential -0.35V (VS.Ag/AgCl) is set as, sensor is successively immersed in above-mentioned solution, and the record time is
400s。
Implement row 9
The present embodiment provides a kind of preparation method of three-dimensional grapheme lactic acid sensor, and the process chart of the method is as schemed
Shown in 1, comprise the following steps that：
1. PECVD is adopted, the Copper Foil of 30 μ m-thicks is lain in a horizontal plane in the glass tubing of growth furnace, successively process intensification,
Annealing, three steps of growth.Condition is respectively：Heat up：50min is warming up to 700 DEG C, hydrogen=5sccm；Annealing：80min, dimension
750 DEG C are held, hydrogen=15sccm；Growth：110min, maintains 650 DEG C, methane:Hydrogen=6：6sccm, radio frequency=400mw；It is raw
Length closes radio frequency and methane after terminating, hydrogen is arranged to 14sccm, opens fan fast cooling, and finally gently taking out growth has
The Copper Foil of Graphene wall.
2. Fe (the NO of 0.5M are prepared3)3Copper etching liquid, then inserts solution by the fritter that Copper Foil is cut to 2.0cm*2.5cm
Surface, to grow facing up of having Graphene wall, overnight fully etching Copper Foil under room temperature condition.Then, Jing plasmas are used
Process the Graphene wall that the 6cm*6cm PET films of (hydrophilic group in exposure pet sheet face, convenient to fit with Graphene wall) will be floated
Pick up, Graphene wall is transferred in ultra-pure water 30min*3 time fully to remove the metal ions such as remaining copper, ferrum.Then,
PET picks up the Graphene wall that cleaned and is put in baking oven and dries under conditions of 120*20min, makes Graphene wall firm with PET
Gu laminating.
3. silk screen print method printed electrode (as shown in Figure 1) is adopted.First Ag/AgCl ink is printed on PET as ginseng
Than electrode, 100 DEG C are toasted 22 minutes；Then, carbon printing ink is printed on as working electrode and to electrode on PET, 95 DEG C of bakings
15-30 18 minutes；Again one layer of insulation paste is printed again in working electrode surface, 85 DEG C are toasted 18 minutes；Finally by three electrodes point
Do not connect conductive filamentary silver and draw circuit.
4. weigh 3.3g shitosans to be dissolved in the acetic acid solution of 100ml 1.0%, room temperature with 250 revs/min stirring 3h with
Prepare chitosan solution；To on GNWs, room temperature is dried to draw 15 μ L Lox solution (concentration is 600IU/mL) drop coatings；Take 10 μ L
3.3% chitosan solution drop coating to GNWs/ enzymes surface, dry by room temperature；The glutaraldehyde drop coating for taking 20 μ L 1.0% is layed onto GNWs/
Enzyme/shitosan surface, room temperature dries 1h；Electrode is immersed in the 0.1M PBSs of pH 7.0,4 DEG C overnight removing electricity
Extremely go up the Lactate Oxidase of remnants；4 DEG C are stored in the 0.1M PBSs of pH7.0.
5. in concentration be 0.1M, the PBS that pH value is 7.0 in compound concentration for 100mM lactic acid solution；In oxygen
In changing-going back virgin curve test process, potential is set as into -1.0V-+1.0V (VS.Ag/AgCl), sensor is successively immersed in
In above-mentioned solution, start recording electric current after 2min, the record time is 60s；In time current curve test process, by potential
It is set as -0.35V (VS.Ag/AgCl), sensor is immersed in above-mentioned solution and records the time for 60s；Survey in product sensitivity
During examination, compound concentration is 0mM, 0.5mM, 50mM, 100mM, 200mM, 400mM in PBS (0.1M, pH7.0) buffer
Lactic acid solution, potential is set as into -0.35V (VS.Ag/AgCl), sensor is successively immersed in above-mentioned solution, during record
Between be 400s.
Claims (1)
1. a kind of preparation method of three-dimensional grapheme lactic acid sensor, it is characterised in that the method step is as follows：
(1) preparation of Graphene wall
Using 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 rising
Temperature, annealing, three steps of growth；Wherein, temperature-rise period is warming up to 500～800 DEG C for 30～90min, and hydrogen=5～
30sccm；Annealing process is 30～90min, maintains 600～800 DEG C, hydrogen=5～15sccm；Growth course be 60～
120min, maintains 600～800 DEG C, methane:Hydrogen=4:8sccm, radio frequency=200～500mw；Growth closes radio frequency after terminating
And methane, hydrogen setting to 2～20sccm, opening fan fast cooling, finally gently taking out growth has the copper of Graphene wall
Paper tinsel；
(2) transfer of Graphene wall
A) compound concentration is the Fe (NO of 0.1M～0.5M3)3Copper etching liquid, b) by Copper Foil be cut into size for 0.2cm*0.2cm～
The fritter that 3cm*3cm is not waited inserts solution surface, is faced up with growing have Graphene wall one, overnight with abundant under room temperature condition
Etching Copper Foil, c) with plasma treated size be 0.2cm*0.2cm～3cm*3cm not wait PET film will float graphite
Alkene wall is picked up, and Graphene wall is transferred to into 30min*3～5 time in ultra-pure water and removes the metal ions such as remaining copper, ferrum with abundant,
D) the Graphene wall that cleaned is picked up with PET and is put in baking oven and dried under conditions of 80～150 DEG C of * 5～60min, make stone
Black alkene wall and PET secure fits；
(3) preparation of electrode
Using silk screen print method printed electrode, a) Ag/AgCl ink is printed on PET as reference electrode, at 60-100 DEG C
Scope toasts 5-25 minutes, b) is printed on carbon printing ink on PET as working electrode and to electrode, in 80-120 DEG C of scope baking
15-30 minutes, c) one layer of insulation paste is printed again in working electrode surface, toasted 5-20 minutes in the range of 50-90 DEG C, d) will
Three electrodes connect respectively conductive filamentary silver and draw circuit；
(4) sensor electrode is surface-functionalized
A) Lactate Oxidase is loaded on three-dimensional grapheme nm wall, room temperature is dried, b) configures 2-20 μ L, concentration is 100-
The chitosan solution of 800IU/ml, c) by 2-20 μ L, concentration is that 0.3-3.5% chitosan solutions drop is layed onto three-dimensional grapheme/breast
Acid oxidase surface, d) is fixed to three-dimensional grapheme wall/enzyme/shitosan surface, room temperature by the glutaraldehyde that concentration is 0.25-2%
A period of time is dried, e) Lactate Oxidase remaining on cleaning electrode and is preserved；It is used during cleaning and preservation
PBS solution concentration is 0.1M, and pH=7.0, the temperature of process is 2-8 DEG C；
(5) lactate detection of sensor electrode
A) the making lactic acid solution in PBS, b) tests transducer sensitivity and tests sensor stability, and c) test is passed
Sensor specificity；Wherein, the lactic acid solution concentration range that the PBS that concentration is 0.1M, pH value is 7.0 is prepared is in 1-
100mM；In oxidation-reduction curve test process, scanning potential interval is set as into -1.0V -+1.0V, reference electrode is
Ag/AgCl, sensor is successively immersed in above-mentioned solution, start recording electric current after 2min, and the record time is 60s；When it is m-
In current curve test process, potential is set as into -0.35V, reference electrode is Ag/AgCl, sensor is immersed in above-mentioned molten
The time is recorded in liquid for 60s；During sensitivity test, electrode is immersed in successively in the lactic acid solution of 0.1-400mM
After 2min, corresponding three electrodes are connected, open electrochemical workstation, potential is set as into -0.35V, reference electrode is Ag/
AgCl, using cyclic voltammetry start recording electric current, the record time is 400s.
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| CN107271517B (en) * | 2017-07-03 | 2019-06-25 | 中国人民解放军第三军医大学 | A kind of sweat lactic acid device for fast detecting |
| CN108468036B (en) * | 2018-02-24 | 2019-12-10 | 东南大学 | preparation method of super-soft semitransparent conductive film |
| CN110907502A (en) * | 2018-09-17 | 2020-03-24 | 中国科学院大连化学物理研究所 | Aniline sensor taking three-dimensional foamy graphene material as sensing film |
| CN111007127A (en) * | 2019-12-24 | 2020-04-14 | 湖南海源医疗科技股份有限公司 | Application of ordered vertical porous graphene in preparation of blood glucose test paper |
| CN113533469B (en) * | 2021-07-13 | 2023-12-29 | 东南大学 | Lactate ion sensor based on graphene/polypyrrole and preparation method and application thereof |
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| CN104887227A (en) * | 2015-04-16 | 2015-09-09 | 中国科学院重庆绿色智能技术研究院 | Graphene flexible surface myoelectricity electrode and preparing method thereof |
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| CN104887227A (en) * | 2015-04-16 | 2015-09-09 | 中国科学院重庆绿色智能技术研究院 | Graphene flexible surface myoelectricity electrode and preparing method thereof |
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