CN108169306A - A kind of preparation method of the high-performance biosensor based on graphene aerogel - Google Patents
A kind of preparation method of the high-performance biosensor based on graphene aerogel Download PDFInfo
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- CN108169306A CN108169306A CN201810170378.4A CN201810170378A CN108169306A CN 108169306 A CN108169306 A CN 108169306A CN 201810170378 A CN201810170378 A CN 201810170378A CN 108169306 A CN108169306 A CN 108169306A
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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 a kind of preparation methods of the high-performance biosensor based on graphene aerogel, it is characterised in that：Include the following steps：（1）The preparation of graphene aerogel/Pt nanoparticle：Graphene oxide powder adds in deionized water, adds in chloroplatinic acid, is encapsulated into polytetrafluoroethylene (PTFE) autoclave, cooled to room temperature after reaction, graphene hydrogel/nano platinum particle made from taking-up.Then use CO2Supercritical extract seasoning obtains graphene aerogel/nano platinum particle.（2）The preparation of enzyme electrode：Take graphene aerogel/nano platinum particle, add in absolute ethyl alcohol, add in biological enzyme, graphene aerogel/nano platinum particle/biology enzyme solutions are made, it is added dropwise in glassy carbon electrode surface, Nafion solution is added dropwise, obtains graphene aerogel/nano platinum particle/biological enzyme glass-carbon electrode, replace the basic structural unit fixed enzyme membrane of conventional biosensor, so as to which a kind of high-performance biosensor based on graphene aerogel will be obtained.
Description
Technical field
The present invention relates to the preparation fields of biosensor, and in particular to a kind of high-performance life based on graphene aerogel
The preparation method of object sensor.
Background technology
Clinical medicine shows that the generation of many diseases of the mankind is all related with the metabolite concentration abnormality in human body, such as urinates
Acid concentration is excessively high to cause ventilation, and cholesterol concentration is excessively high to cause cardiovascular and cerebrovascular disease etc..Have to effectively prevent these
Disease caused by body metabolism substance concentration abnormality, people need a kind of simple and effective detection method, and existing medicine is examined
Survey processing procedures for needing expensive instrument and equipment, the operation of profession and complexity etc. more.Common biology passes on the market at present
Sensor there are sensitivity it is low, poor specificity, be disturbed the shortcomings of ability is low, response speed is slow, so novel high-performance biology pass
The preparation of sensor becomes the emphasis of research concern.
Invention content
In order to solve the deficiencies in the prior art, the present invention provides a kind of detection range is wide, high sensitivity, the response time
Short, the low high-performance biosensor based on graphene aerogel of detectable limit preparation method.Include the following steps：
（1）It is prepared by graphene oxide：Using the concentrated sulfuric acid of more than 98% volume fraction of 25ml ~ 100ml, 1g ~ 2g nitric acid is added in
Potassium, ice bath stirring add in 2g graphite ice bath stirring 30min ~ 60min, be slowly added to the potassium permanganate solution 20g of 50wt% ~
30g, ice bath stirring 30min ~ 60min are transferred to 20 DEG C ~ 30 DEG C stirring in water bath 30min ~ 60min, are slowly added to 200ml deionizations
Water is transferred to 90 DEG C ~ 110 DEG C stirring in water bath 10min ~ 30min, is then slowly added into the 20% ~ 30% of 500ml deionized waters and 40ml
Hydrogen peroxide stirring 10min.Until the color of mixture becomes glassy yellow, by mixture centrifuge washing to neutrality, it is dry after i.e.
Obtain graphene oxide powder.
（2）The preparation of graphene hydrogel/nano platinum particle：100mg-300mg graphene oxide powders are taken, are added in
30ml-100ml deionized waters, using ultrasonic stirrer ultrasonic agitation 0.5h or so, then by a solution heating water bath 0.5h left sides
The right side finally adds in the deionized water of 20ml-50ml, and 0.5h-0.8h is stirred by ultrasonic with ultrasonic stirrer again, you can obtains
The graphene solution that graphene oxide uniformly disperses.
It takes and prepares a concentration of 30-50% platinum acid chloride solutions 50-150ml, add in prepared graphene solution in three times
In, and 2h is persistently stirred with magnetic stirring apparatus, obtain mixed solution.Mixed solution obtained is encapsulated into polytetrafluoroethylene (PTFE) high pressure
In reaction kettle, 10h-12h, cooled to room temperature after reaction, graphite made from taking-up are reacted under conditions of 180 DEG C -200 DEG C
Alkene hydrogel/nano platinum particle.
Using soaked in absolute ethyl alcohol cleaning graphene hydrogel/nano platinum particle, and use ultrasonic echography 0.5h.Then
Use CO2Supercritical extract seasoning dry pure graphene hydrogel/nano platinum particle under the conditions of 10MPa, 45 DEG C, with
Graphene aerogel/nano platinum particle is obtained, by graphene aerogel/nano platinum particle under conditions of 1000 DEG C -1200 DEG C
3h-5h is heated, has obtained pure graphene aerogel/nano platinum particle.
（3）The preparation of enzyme electrode：Glass-carbon electrode is taken, electrode is ground using sand paper, polishing powder, later using anhydrous
Ethyl alcohol cleaning impregnates 5min-15min, then with deionized water ultrasound 5min-15min, to obtain the glass-carbon electrode of pure qualification.
In the environment of 4 DEG C -6 DEG C, 10mg-50mg graphene aerogels/nano platinum particle is taken, adds in 300ml-500ml
Absolute ethyl alcohol, using ultrasonic stirrer ultrasound 0.5h, add in 10mg-20mg biological enzyme later, reuse ultrasonic wave and stir
Device ultrasound 0.5h is mixed, graphene aerogel/nano platinum particle/biology enzyme solutions are made.
Graphene aerogel/nano platinum particle/biology enzyme solutions dropwise addition of 5 μ l-10 μ l is taken in the glass carbon electricity of pure qualification
Electrode is placed in 4 DEG C of -6 DEG C of isoperibols drying 10h-12h, the Nafion solution that 3 μ l have been configured is taken to be added dropwise in electrode by pole surface
Electrode is placed in 4 DEG C of -6 DEG C of isoperibols drying 4h-5h again, obtains graphene aerogel/nano platinum particle/biology by surface
The glass-carbon electrode of enzyme modification.
Using the glass-carbon electrode of graphene aerogel/nano platinum particle/biology enzyme modification, replace conventional biosensor
Basic structural unit fixed enzyme membrane, so as to which a kind of high-performance biosensor based on graphene aerogel will be obtained.
Specific embodiment
It is described in detail to various aspects of the present invention below, unless specific instructions, various raw materials of the invention can pass through
It is prepared or is commercially available according to the conventional method of this field.
Embodiment 1
（1）It is prepared by graphene oxide：Using the concentrated sulfuric acid of more than 98% volume fraction of 25ml, 1g potassium nitrate is added in, ice bath stirs
It mixes, adds in 2g graphite ice bath stirring 30min, be slowly added to potassium permanganate solution 20g, the ice bath stirring 30min of 50wt%, turn
Enter 20 DEG C of stirring in water bath 30min, be slowly added to 200ml deionized waters, be transferred to 90 DEG C of stirring in water bath 10min, be then slowly added into
20% hydrogen peroxide of 500ml deionized waters and 40ml stir 10min.Until the color of mixture becomes glassy yellow, by mixture
Centrifuge washing is to neutrality, up to graphene oxide powder after drying.
（2）The preparation of graphene hydrogel/nano platinum particle：100mg graphene oxide powders are taken, add in 30ml deionizations
Water using ultrasonic stirrer ultrasonic agitation 0.5h or so, then by solution heating water bath 0.5h or so, finally adds in again
The deionized water of 20ml, and 0.5h is stirred by ultrasonic with ultrasonic stirrer, you can obtain the graphite that graphene oxide uniformly disperses
Alkene solution.
It takes and prepares a concentration of 30% platinum acid chloride solution 50ml, add in prepared graphene solution, be used in combination in three times
Magnetic stirring apparatus persistently stirs 2h, obtains mixed solution.Mixed solution obtained is encapsulated into polytetrafluoroethylene (PTFE) autoclave
It is interior, react 10h under conditions of 180 DEG C -200 DEG C, cooled to room temperature after reaction, graphene hydrogel made from taking-up/
Nano platinum particle.
Using soaked in absolute ethyl alcohol cleaning graphene hydrogel/nano platinum particle, and use ultrasonic echography 0.5h.Then
Use CO2Supercritical extract seasoning dry pure graphene hydrogel/nano platinum particle under the conditions of 10MPa, 45 DEG C, with
Graphene aerogel/nano platinum particle is obtained, graphene aerogel/nano platinum particle is heated into 3h under conditions of 1000 DEG C,
Pure graphene aerogel/nano platinum particle is obtained.
（3）The preparation of enzyme electrode：Glass-carbon electrode is taken, electrode is ground using sand paper, polishing powder, later using anhydrous
Ethyl alcohol cleaning impregnates 5min, then with deionized water ultrasound 5min, to obtain the glass-carbon electrode of pure qualification.
In the environment of 4 DEG C -6 DEG C, 10mg graphene aerogels/nano platinum particle is taken, adds in the absolute ethyl alcohol of 300ml,
Using ultrasonic stirrer ultrasound 0.5h, 10mg biological enzyme is added in later, reuses ultrasonic stirrer ultrasound 0.5h, be made
Graphene aerogel/nano platinum particle/biology enzyme solutions.
Graphene aerogel/nano platinum particle/biology enzyme solutions dropwise addition of 5 μ l is taken in the glass-carbon electrode table of pure qualification
Electrode is placed in 4 DEG C of -6 DEG C of isoperibols drying 10h, the Nafion solution that 3 μ l have been configured is taken to be added dropwise in electrode surface by face, then
It is secondary that electrode is placed in 4 DEG C of -6 DEG C of isoperibols drying 4h, obtain the glass of graphene aerogel/nano platinum particle/biology enzyme modification
Carbon electrode.
Using the glass-carbon electrode of graphene aerogel/nano platinum particle/biology enzyme modification, replace conventional biosensor
Basic structural unit fixed enzyme membrane, so as to which a kind of high-performance biosensor based on graphene aerogel will be obtained.
Embodiment 2
（1）It is prepared by graphene oxide：Using the concentrated sulfuric acid of more than 98% volume fraction of 50ml, 1.5g potassium nitrate is added in, ice bath stirs
It mixes, adds in 2g graphite ice bath stirring 45min, be slowly added to potassium permanganate solution 25g, the ice bath stirring 45min of 50wt%, turn
Enter 25 DEG C of stirring in water bath 45min, be slowly added to 200ml deionized waters, be transferred to 100 DEG C of stirring in water bath 20min, be then slowly added into
25% hydrogen peroxide of 500ml deionized waters and 40ml stir 10min.Until the color of mixture becomes glassy yellow, by mixture
Centrifuge washing is to neutrality, up to graphene oxide powder after drying.
（2）The preparation of graphene hydrogel/nano platinum particle：Take 200mg graphene oxide powders, add in 100ml go from
Sub- water using ultrasonic stirrer ultrasonic agitation 0.5h or so, then by solution heating water bath 0.5h or so, finally adds in again
The deionized water of 20ml, and 0.8h is stirred by ultrasonic with ultrasonic stirrer, you can obtain the graphite that graphene oxide uniformly disperses
Alkene solution.
It takes and prepares a concentration of 40% platinum acid chloride solution 100ml, add in three times in prepared graphene solution, and
2h is persistently stirred with magnetic stirring apparatus, obtains mixed solution.Mixed solution obtained is encapsulated into polytetrafluoroethylene (PTFE) reaction under high pressure
In kettle, 12h, cooled to room temperature after reaction, graphene water-setting made from taking-up are reacted under conditions of 180 DEG C -200 DEG C
Glue/nano platinum particle.
Using soaked in absolute ethyl alcohol cleaning graphene hydrogel/nano platinum particle, and use ultrasonic echography 0.5h.Then
Use CO2Supercritical extract seasoning dry pure graphene hydrogel/nano platinum particle under the conditions of 10MPa, 45 DEG C, with
Graphene aerogel/nano platinum particle is obtained, graphene aerogel/nano platinum particle is heated into 3h under conditions of 1200 DEG C,
Pure graphene aerogel/nano platinum particle is obtained.
（3）The preparation of enzyme electrode：Glass-carbon electrode is taken, electrode is ground using sand paper, polishing powder, later using anhydrous
Ethyl alcohol cleaning impregnates 10min, then with deionized water ultrasound 10min, to obtain the glass-carbon electrode of pure qualification.
In the environment of 4 DEG C -6 DEG C, 30mg graphene aerogels/nano platinum particle is taken, adds in the absolute ethyl alcohol of 300ml,
Using ultrasonic stirrer ultrasound 0.5h, 10mg biological enzyme is added in later, reuses ultrasonic stirrer ultrasound 0.5h, be made
Graphene aerogel/nano platinum particle/biology enzyme solutions.
Graphene aerogel/nano platinum particle/biology enzyme solutions dropwise addition of 10 μ l is taken in the glass-carbon electrode table of pure qualification
Electrode is placed in 4 DEG C of -6 DEG C of isoperibols drying 12h, the Nafion solution that 3 μ l have been configured is taken to be added dropwise in electrode surface by face, then
It is secondary that electrode is placed in 4 DEG C of -6 DEG C of isoperibols drying 5h, obtain the glass of graphene aerogel/nano platinum particle/biology enzyme modification
Carbon electrode.
Using the glass-carbon electrode of graphene aerogel/nano platinum particle/biology enzyme modification, replace conventional biosensor
Basic structural unit fixed enzyme membrane, so as to which a kind of high-performance biosensor based on graphene aerogel will be obtained.
Embodiment 3
（1）It is prepared by graphene oxide：Using the concentrated sulfuric acid of more than 98% volume fraction of 100ml, 2g potassium nitrate is added in, ice bath stirs
It mixes, adds in 2g graphite ice bath stirring 60min, be slowly added to potassium permanganate solution 30g, the ice bath stirring 60min of 50wt%, turn
Enter 30 DEG C of stirring in water bath 60min, be slowly added to 200ml deionized waters, be transferred to 110 DEG C of stirring in water bath 30min, be then slowly added into
30% hydrogen peroxide of 500ml deionized waters and 40ml stir 10min.Until the color of mixture becomes glassy yellow, by mixture
Centrifuge washing is to neutrality, up to graphene oxide powder after drying.
（2）The preparation of graphene hydrogel/nano platinum particle：Take 300mg graphene oxide powders, add in 100ml go from
Sub- water using ultrasonic stirrer ultrasonic agitation 0.5h or so, then by solution heating water bath 0.5h or so, finally adds in again
The deionized water of 50ml, and 0.8h is stirred by ultrasonic with ultrasonic stirrer, you can obtain the graphite that graphene oxide uniformly disperses
Alkene solution.
It takes and prepares a concentration of 50% platinum acid chloride solution 150ml, add in three times in prepared graphene solution, and
2h is persistently stirred with magnetic stirring apparatus, obtains mixed solution.Mixed solution obtained is encapsulated into polytetrafluoroethylene (PTFE) reaction under high pressure
In kettle, 12h, cooled to room temperature after reaction, graphene water-setting made from taking-up are reacted under conditions of 180 DEG C -200 DEG C
Glue/nano platinum particle.
Using soaked in absolute ethyl alcohol cleaning graphene hydrogel/nano platinum particle, and use ultrasonic echography 0.5h.Then
Use CO2Supercritical extract seasoning dry pure graphene hydrogel/nano platinum particle under the conditions of 10MPa, 45 DEG C, with
Graphene aerogel/nano platinum particle is obtained, graphene aerogel/nano platinum particle is heated into 5h under conditions of 1200 DEG C,
Pure graphene aerogel/nano platinum particle is obtained.
（3）The preparation of enzyme electrode：Glass-carbon electrode is taken, electrode is ground using sand paper, polishing powder, later using anhydrous
Ethyl alcohol cleaning impregnates 15min, then with deionized water ultrasound 15min, to obtain the glass-carbon electrode of pure qualification.
In the environment of 4 DEG C -6 DEG C, 50mg graphene aerogels/nano platinum particle is taken, adds in the absolute ethyl alcohol of 500ml,
Using ultrasonic stirrer ultrasound 0.5h, 20mg biological enzyme is added in later, reuses ultrasonic stirrer ultrasound 0.5h, be made
Graphene aerogel/nano platinum particle/biology enzyme solutions.
Graphene aerogel/nano platinum particle/biology enzyme solutions dropwise addition of 10 μ l is taken in the glass-carbon electrode table of pure qualification
Electrode is placed in 4 DEG C of -6 DEG C of isoperibols drying 12h, the Nafion solution that 3 μ l have been configured is taken to be added dropwise in electrode surface by face, then
It is secondary that electrode is placed in 4 DEG C of -6 DEG C of isoperibols drying 5h, obtain the glass of graphene aerogel/nano platinum particle/biology enzyme modification
Carbon electrode.
Using the glass-carbon electrode of graphene aerogel/nano platinum particle/biology enzyme modification, replace conventional biosensor
Basic structural unit fixed enzyme membrane, so as to which a kind of high-performance biosensor based on graphene aerogel will be obtained.
The present invention is not limited to the embodiment, anyone should learn that the structure made under the enlightenment of the present invention becomes
Change, the technical schemes that are same or similar to the present invention are each fallen within protection scope of the present invention.
The present invention be not described in detail technology, shape, construction part be known technology.
Claims (5)
1. a kind of preparation method of the high-performance biosensor based on graphene aerogel, it is characterised in that：Take 5 μ l-10 μ l
Graphene aerogel/nano platinum particle/biology enzyme solutions glassy carbon electrode surface in pure qualification is added dropwise, electrode is placed in 4
DEG C of -6 DEG C isoperibols drying 10h-12h take the Nafion solution dropwise addition that 3 μ l have been configured again to put electrode in electrode surface
4h-5h is dried in 4 DEG C of -6 DEG C of isoperibols, obtains the glass-carbon electrode of graphene aerogel/nano platinum particle/biology enzyme modification.
Using the glass-carbon electrode of graphene aerogel/nano platinum particle/biology enzyme modification, replace the basic structure of conventional biosensor
Unit fixed enzyme membrane, so as to which a kind of high-performance biosensor based on graphene aerogel will be obtained.
2. a kind of preparation method of high-performance biosensor based on graphene aerogel according to claim 1,
It is characterized in that：Graphene aerogel/nano platinum particle/biology the enzyme solutions are prepared according to the following steps, in 4 DEG C -6 DEG C of environment
Under, 10mg-50mg graphene aerogels/nano platinum particle is taken, the absolute ethyl alcohol of 300ml-500ml is added in, is stirred using ultrasonic wave
Device ultrasound 0.5h is mixed, 10mg-20mg biological enzyme is added in later, reuses ultrasonic stirrer ultrasound 0.5h, graphene is made
Aeroge/nano platinum particle/biology enzyme solutions.
3. a kind of preparation method of high-performance biosensor based on graphene aerogel according to claim 1,
It is characterized in that：The glass-carbon electrode of the pure qualification is prepared according to the following steps, glass-carbon electrode is taken, using sand paper, polishing powder to electricity
Pole is ground, later using washes of absolute alcohol, impregnate 5min-15min, then with deionized water ultrasound 5min-15min, with
Obtain the glass-carbon electrode of pure qualification.
4. a kind of preparation method of high-performance biosensor based on graphene aerogel according to claim 2,
It is characterized in that：Graphene aerogel/the nano platinum particle is prepared according to the following steps, takes 100mg-300mg graphene oxide powder
End adds in 30ml-100ml deionized waters, using ultrasonic stirrer ultrasonic agitation 0.5h or so, then by solution heating water bath
0.5h or so finally adds in the deionized water of 20ml-50ml, and 0.5h-0.8h is stirred by ultrasonic with ultrasonic stirrer again, i.e.,
The graphene solution that graphene oxide uniformly disperses is can obtain, takes and prepares a concentration of 30-50% platinum acid chloride solutions 50-150ml,
It adds in three times in prepared graphene solution, and 2h is persistently stirred with magnetic stirring apparatus, obtain mixed solution.It will be made
Mixed solution be encapsulated into polytetrafluoroethylene (PTFE) autoclave, 10h-12h is reacted under conditions of 180 DEG C -200 DEG C, react
Cooled to room temperature afterwards, graphene hydrogel/nano platinum particle made from taking-up clean graphene using soaked in absolute ethyl alcohol
Hydrogel/nano platinum particle, and use ultrasonic echography 0.5h.Then use CO2Supercritical extract seasoning is in 10MPa, and 45
Dry pure graphene hydrogel/nano platinum particle under the conditions of DEG C, to obtain graphene aerogel/nano platinum particle, by stone
Black alkene aeroge/nano platinum particle heats 3h-5h under conditions of 1000 DEG C -1200 DEG C, has obtained pure graphene airsetting
Glue/nano platinum particle.
5. a kind of preparation method of high-performance biosensor based on graphene aerogel according to claim 4,
It is characterized in that：The graphene oxide powder is prepared according to the following steps, using more than 98% volume fraction of 25ml~100ml
The concentrated sulfuric acid, adds in 1g~2g potassium nitrate, and ice bath stirring adds in 2g graphite ice bath stirring 30min~60min, is slowly added to
Potassium permanganate solution 20g~30g of 50wt%, ice bath stirring 30min~60min are transferred to 20 DEG C~30 DEG C stirring in water bath
30min~60min is slowly added to 200ml deionized waters, is transferred to 90 DEG C~110 DEG C stirring in water bath 10min~30min, Ran Houhuan
Slow 20%~30% hydrogen peroxide stirring 10min for adding in 500ml deionized waters and 40ml.Until the color of mixture becomes bright
Yellow, by mixture centrifuge washing to neutrality, it is dry after up to graphene oxide powder.
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Cited By (1)
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| CN109521072A (en) * | 2018-12-31 | 2019-03-26 | 合肥工业大学 | Detect heteroauxin and salicylic method and its electrochemical sensor used |
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