CN103616422A - Preparation method of graphene/hemoglobin composite film modified electrode - Google Patents
Preparation method of graphene/hemoglobin composite film modified electrode Download PDFInfo
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- CN103616422A CN103616422A CN201310643584.XA CN201310643584A CN103616422A CN 103616422 A CN103616422 A CN 103616422A CN 201310643584 A CN201310643584 A CN 201310643584A CN 103616422 A CN103616422 A CN 103616422A
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Abstract
The invention relates to a preparation method of a graphene/hemoglobin composite film based on a layer by layer self-assembly technology, which belongs to the technical field of preparation of composite material modified electrodes. The preparation method provided by the invention comprises the following steps: soaking a treated electrode in modified graphene aqueous dispersion for a period of time, taking out the electrode, cleaning the electrode through deionized water, then, soaking the electrode in a hemoglobin aqueous solution for a period of time, taking out and cleaning the electrode, and repeating the process to form a uniform graphene/hemoglobin film having a regular layered structure on the surface of the electrode. The graphene/hemoglobin modified electrode prepared by the method is controllable in structural performance, good in sensitivity, reproducibility and stability, wide in linear detection range, low in lower detection limit, simple in preparation process, low in cost and easy to operate.
Description
Technical field:
The invention belongs to based on compound substance modified electrode preparation field, be characterized in adopting self-assembling method layer by layer to prepare Graphene/haemoglobin multi-layer compound film, and use it for the preparation of modified electrode.
Background technology:
In recent years, compound substance modified electrode is a focus of sensor field, is more and more subject to people and payes attention to.
Haemoglobin is a kind of protein that protoheme is prothetic group of take, and the class tetramer consisting of two αHe Liangge β subunits is being born and in biosome, transmitted electronics, transmission oxygen, decomposing H
2o
2deng the vital role relevant with the metabolism of biosome self-energy.Because haemoglobin has the catalytic action to peroxidase and cytochrome, and it is cheap and easy to get, therefore thinks a kind of ideal model with heme molecule protein.Yet, due to the huge spatial configuration of molecules of haemoglobin, its electric activity center is coated in polypeptied chain, again because haemoglobin is by electrode surface strong adsorption, this just causes electrode passivation, and this makes haemoglobin very slow in the transfer rate of electrode surface, and current responsing signal is too little.Therefore, if can introduce certain high conductivity material at electrode surface, and utilize this material to carry out immobilized haemoglobin, this just can effectively solve the problem that haemoglobin can not get watt current response, and can make the biologically active of haemoglobin be retained.In recent years, people utilize promoter effect or electron mediator to improve the transmission of haemoglobin electronics, thereby realize it in the catalytic action of electrode surface.But, the introducing meeting of part promoter produces certain interference to the catalytic performance of haemoglobin, this just exerts an influence to the selective control of sensor, and difficulty and the bad control of speed that for example, in electron mediator (enzyme classes), electronics shifts, and there is the problems such as amboceptor loss, electrode fouling, also can have influence on the control to sensor catalytic process, so most important to selecting of electron mediator.
Graphene is owing to having the performances such as unique structure and excellent electricity, calorifics, mechanics, since 2004, successfully prepared, be a study hotspot in worldwide always, become the advanced special type function material such as electronic material that a class is new, membraneous material, energy storage material, liquid crystal material, catalysis material.Graphene composite material is the key areas of Graphene application, utilizes the good characteristic of Graphene and other Material cladding can give the character of material excellence, for it has opened up more wide application prospect.Compound between Graphene and biomacromolecule shows excellent performance at aspects such as conductance, biologically active and mechanical propertys.Yet, because Graphene chemical stability is high, its surface is inert condition, with other medium interactions a little less than, and between graphene film and sheet, there is stronger Van der Waals force, easily produce and reunite, make it be insoluble in water and conventional organic solvent, bad dispersibility, this has limited further research and the application of graphene composite material greatly.Therefore, further improve and the nature and function of expanding Graphene and compound substance thereof are extremely important.
Summary of the invention:
Problem to be solved by this invention is to provide a kind of preparation method based on Graphene/haemoglobin laminated film modified electrode.It is module units that the method be take respectively Graphene and haemoglobin, by electrostatic interaction, in electrode surface self assembly layer by layer, prepares Graphene/haemoglobin laminated film.Graphene/modified hemoglobin electrode structure performance prepared by the method is controlled, has good sensitivity, reappearance and stability, has wider linear detection range and lower detection lower limit.
The preparation method who the present invention is based on a kind of Graphene/haemoglobin laminated film modified electrode, comprising:
(1) surface after modification being configured to concentration range with carboxyl or sulfonic Graphene is 0.2～10mg/mL, the aqueous dispersions that pH scope is 4～14.It is 0.1～30mg/mL that haemoglobin is configured to concentration range, the aqueous solution that pH scope is 4～6.8.
(2) electrode after processing is immersed in to 10～30min in haemoglobin aqueous solution, then uses a large amount of deionized water rinsings, remove the haemoglobin of electrode surface physisorption, nitrogen dries up, the electrode surface lotus positive electricity obtaining.
(3) electrode after step (2) is processed is immersed in to 10～30min in Graphene aqueous dispersions, then uses a large amount of deionized water rinsings, remove the Graphene of electrode surface physisorption, nitrogen dries up, the electrode surface bear electricity obtaining.
(4) repeating step (2) and (3), obtain Graphene/haemoglobin laminated film modified electrode.
(5) Graphene in above-mentioned steps (1) is after surface modification, and by regulating pH (4～14) value to make its surperficial bear electricity, and energy stable dispersion is in water.And lotus positive electricity in the aqueous dispersions that haemoglobin is 4～6.8 at pH, two kinds of module units of Graphene and haemoglobin can rely on together with electrostatic interaction is assembled into.Graphene aqueous dispersions concentration is 0.2～10mg/mL, and the concentration of haemoglobin aqueous solution is 0.1～30mg/mL.
(6) the present invention compares with existing invention, its remarkable advantage has: (1) has overcome the resulting Graphene class of prior art compound substance bad dispersibility, be difficult for machine-shaping, to function element below construct and performance study causes the problem of very big difficulty; (2) this technology can well be controlled the component concentration of Graphene in compound substance and haemoglobin, arrangement mode and Compound Degree, thereby can be controlled the performance of compound substance, can supermolecule level, to compound substance, design and controlled preparation; (3) autonomous packing technique cost is low layer by layer, and mild condition is simple to operate; (4) Graphene/modified hemoglobin electrode fabrication process that prepared by this technology is simple, and cost is low, easy to operate, has good sensitivity, reappearance and stability, has wider linear detection range and lower detection lower limit.
Accompanying drawing explanation
Fig. 1 is the uv-visible absorption spectra of modified graphene/haemoglobin laminated film.
Embodiment
In order to explain better, below in conjunction with embodiment, the present invention is further explained in detail the present invention.
Embodiment 1
The first step adds 30mg graphene oxide in 250mL flask, adds DMF/H
2the solution 50mL of O (V:V=9:1), stirs ultrasonic processing 30min.In flask, add 0.0248g NaBH
4, 80 ℃ of oil baths 4 hours, drip 10g acrylic acid and 40mL H after cool to room temperature
2o, stirs 30min.Pass into high-purity N
230min, slowly drips 100mg (NH wherein
4)
2s
2o
8(being dissolved in 80mL deionized water), ultrasonic, 60 ℃ of oil baths, stir.After 48h, cool to room temperature, uses 200mLH
2after O dilution, ultrasonic 1h, centrifugal, obtain the Graphene that polyacrylic acid is modified.
Second step, is dissolved in 20mg haemoglobin in 10mL water, obtains the haemoglobin aqueous solution of 2mg/mL.
The 3rd step, is immersed in 20min in haemoglobin aqueous solution by the glass-carbon electrode after processing, and then uses a large amount of deionized water rinsings, and nitrogen dries up, the glass-carbon electrode surface lotus positive electricity obtaining.
The 4th step, is immersed in 20min in Graphene aqueous dispersions by above-mentioned glass-carbon electrode, then uses a large amount of deionized water rinsings, and nitrogen dries up, the glass-carbon electrode surface bear electricity obtaining.
The 5th step, repeats the 3rd step and the 4th step 5 times.
Embodiment 2
The first step adds 50mg graphene oxide in 250mL flask, adds DMF/H
2the solution 80mL of O (V:V=9:1), stirs ultrasonic processing 30min.In flask, add 0.0248gNaBH
4, 80 ℃ of oil baths 4 hours, drip 20g benzene sulfonic acid and 60mL H after cool to room temperature
2o, stirs 30min.Pass into high-purity N
230min, slowly drips 0.2g (NH wherein
4)
2s
2o
8, ultrasonic, 60 ℃ of oil baths, stir.After 48h, cool to room temperature, uses 300mL H
2after O dilution, ultrasonic 1h, centrifugal, obtain the Graphene that polyphenyl sulfonic acid is modified.
Second step, is dissolved in 10mg haemoglobin in 10mL water, obtains the haemoglobin aqueous solution of 1mg/mL.
The 3rd step, is immersed in 20min in haemoglobin aqueous solution by the gold electrode after processing, and then uses a large amount of deionized water rinsings, and nitrogen dries up, the gold electrode surfaces lotus positive electricity obtaining.
The 4th step, is immersed in 20min in Graphene aqueous dispersions by above-mentioned gold electrode, then uses a large amount of deionized water rinsings, and nitrogen dries up, the gold electrode surfaces bear electricity obtaining.
The 5th step, repeats the 3rd step and the 4th step 5 times.
Embodiment 3
The first step adds 50mg graphene oxide in 250mL flask, adds DMF/H
2the solution 80mL of O (V:V=9:1), stirs ultrasonic processing 30min.In flask, add 0.0248g NaBH
4, 80 ℃ of oil baths 4 hours, drip 20g acrylic acid and 60mL H after cool to room temperature
2o, stirs 30min.Pass into high-purity N
230min, slowly drips 0.2g (NH wherein
4)
2s
2o
8, ultrasonic, 60 ℃ of oil baths, stir.After 48h, cool to room temperature, uses 300mLH
2after O dilution, ultrasonic 1h, centrifugal, obtain the Graphene that polyphenyl sulfonic acid is modified.
Second step, is dissolved in 10mg haemoglobin in 10mL water, obtains the haemoglobin aqueous solution of 1mg/mL.
The 3rd step, is immersed in 30min in haemoglobin aqueous solution by the ITO electrode after processing, and then uses a large amount of deionized water rinsings, and nitrogen dries up, the ITO electrode surface lotus positive electricity obtaining.
The 4th step, is immersed in 30min in Graphene aqueous dispersions by above-mentioned ITO electrode, then uses a large amount of deionized water rinsings, and nitrogen dries up, the ITO electrode surface bear electricity obtaining.
The 5th step, repeats the 3rd step and the 4th step 5 times.
Above-described embodiment is used for the present invention that explains, rather than limits the invention, and in the protection domain of spirit of the present invention and claim, any modification and change that the present invention is made, all fall into protection scope of the present invention.
Claims (5)
1. the preparation method based on Graphene/haemoglobin laminated film modified electrode, its making step is:
(1) configuration Graphene aqueous dispersions and haemoglobin aqueous solution;
(2) the electronegative electrode in the surface with after piranha agent treated is immersed in to 10～30min in haemoglobin aqueous solution, then uses a large amount of deionized water rinsings, nitrogen dries up;
(3) electrode after step (2) is processed is immersed in to 10～30min in Graphene aqueous dispersions, then uses a large amount of deionized water rinsings, nitrogen dries up;
(4) repeating step (2) and (3) n time (n=4～30), obtain based on Graphene/haemoglobin laminated film modified electrode.
2. a kind of preparation method based on Graphene/haemoglobin laminated film modified electrode according to claim 1, it is characterized in that Graphene used in step (1) is for process surface modification is afterwards with sulfonic group or carboxyl, it has good water dispersible and stability.
3. a kind of preparation method based on Graphene/haemoglobin laminated film modified electrode according to claim 1, it is characterized in that in step (1), Graphene aqueous dispersions concentration is 0.2～10mg/mL, the concentration of haemoglobin aqueous solution is 0.1～30mg/mL.
4. a kind of preparation method based on Graphene/haemoglobin laminated film modified electrode according to claim 1, the pH that it is characterized in that Graphene aqueous dispersions is 4～14, the pH of haemoglobin aqueous solution is 4～6.8.
5. a kind of preparation method based on Graphene/haemoglobin laminated film modified electrode according to claim 1, is characterized in that substrate is to process glass-carbon electrode later, gold electrode, ITO electrode etc.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105403606A (en) * | 2015-11-10 | 2016-03-16 | 西安建筑科技大学 | Preparation method for carbon cloth electrode based on cobalt phosphide/hemoglobin modification |
| CN107037104A (en) * | 2017-05-15 | 2017-08-11 | 海南师范大学 | It is a kind of based on the protein electrochemistry senser element of modified by graphene quantum dot electrode and its preparation and application |
| CN108241012A (en) * | 2016-12-27 | 2018-07-03 | 中国科学院金属研究所 | The preparation method and applications of hemoglobin sensor electrode based on graphene |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105403606A (en) * | 2015-11-10 | 2016-03-16 | 西安建筑科技大学 | Preparation method for carbon cloth electrode based on cobalt phosphide/hemoglobin modification |
| CN108241012A (en) * | 2016-12-27 | 2018-07-03 | 中国科学院金属研究所 | The preparation method and applications of hemoglobin sensor electrode based on graphene |
| CN107037104A (en) * | 2017-05-15 | 2017-08-11 | 海南师范大学 | It is a kind of based on the protein electrochemistry senser element of modified by graphene quantum dot electrode and its preparation and application |
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Application publication date: 20140305 |