CN108614022A - A kind of electrochemical method of carbon paper-gold nano grain combination electrode detection NADH - Google Patents
A kind of electrochemical method of carbon paper-gold nano grain combination electrode detection NADH Download PDFInfo
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- CN108614022A CN108614022A CN201810565299.3A CN201810565299A CN108614022A CN 108614022 A CN108614022 A CN 108614022A CN 201810565299 A CN201810565299 A CN 201810565299A CN 108614022 A CN108614022 A CN 108614022A
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- 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
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- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
- G01N27/3277—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction being a redox reaction, e.g. detection by cyclic voltammetry
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- 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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Abstract
The invention belongs to electrochemical analysis detection fields, disclose a kind of electrochemical method of carbon paper gold nano grain combination electrode detection NADH.The preparation method of the combination electrode used for, using cyclic voltammetry by gold nano grain be deposited on carbon paper at least 1 enclose, you can obtain.The advantages of combination electrode that the present invention uses：Have the characteristics that prepare that simple, of low cost, disposable, sensitivity is strong, stability is good, reproducibility is high.And context of detection is quantified with vast potential for future development in NADH.
Description
Technical field
The invention belongs to electrochemical sensing fields, and in particular to a kind of novel carbon paper-gold nano grain combination electrode (CP-
AuNxE the electrochemical method of NADH) is detected.
Background technology
Nicotinamide adenine dinucleotide (β-Nicotinamide adenine dinucleotide, NADH) and its oxygen
Change form (NAD+), it is the important coenzyme needed for human life activity, directly participates in the generation of energy and consumption process in cell.
The shortage of NADH easily causes tired, absent minded, brain response is blunt etc. in human body, serious to also result in A Erci
The silent disease in sea and parkinsonism.And content excessively also results in the symptoms such as insomnia, depression.Therefore, NADH is quick, sensitive, accurate
Assay method plays a crucial role clinical diagnosis and pharmaceuticals industry.
Currently, capillary electrophoresis, chemoluminescence method, high performance liquid chromatography and electrochemical method can be used for examining
Survey NADH.Wherein, different from other several traditional detection methods, electrochemical detection method do not need expensive instrument and
Complicated operation etc..The characteristics of Electrochemical Detection is easy to operate, high sensitivity, detection limit is low, at low cost, it is miniature to be easily achieved
Change etc., by global extensive concern.The bottleneck currently encountered is：First, anode detections of the NADH on ordinary electrode
Need larger overpotential；Secondly, the oxidation product that reaction process generates, which is easy to be adsorbed onto electrode surface, causes sensor passivation；
Again, its active material of traditional modified electrode is easy to fall off or occurs agglomeration in electrode surface, so that detection NADH
It is ineffective.Moreover, before preparing traditional modified electrode is wasted to the preprocessing process of glass-carbon electrode or gold electrode many times,
It is unfavorable for the practical application of the sensing platform.
Invention content
In order to solve the deficiencies in the prior art, an object of the present invention is to provide a kind of carbon paper-for detecting NADH
The preparation method of gold nano grain combination electrode, electrode prepared by this method can realize the highly sensitive detection of NADH.
To achieve the goals above, the technical scheme is that：
A kind of preparation method for detecting carbon paper-gold nano grain combination electrode of NADH, using electrochemical deposition method
Gold nano grain is deposited on carbon paper at least 1 to enclose, you can obtain carbon paper-gold nano grain combination electrode for detecting NADH.
Combination electrode prepared by the present invention, stability is high, and preparation process is simple, and can realize the highly sensitive inspection of NADH
It surveys.
The second object of the present invention is to provide a kind of carbon paper-gold nano for detecting NADH prepared by above-mentioned preparation method
Particles dispersed electrode.
The third object of the present invention is to provide a kind of electrochemical method of detection NADH, using above-mentioned electrode as working electrode,
The solution containing NADH is detected using cyclic voltammetry (CV) or differential pulse voltammetry (DPV).
Combination electrode provided by the invention can be obtained is beneficial to the reason of detection result：A) .Au nano-particles have good
Good electric conductivity and quite high current density so that transmission speed of the electronics on CP-Au electrodes is accelerated；B) .Au nanometers
Successful growth of the particle in carbon paper substrate makes CP-AuNxE possess great specific surface area, exposes more active sites
Point；C) .Au nano-particles itself enhance the electrode to NADH's to the catalytic performance and Au of NADH and the synergistic effect of CP
Catalytic oxidation performance.
Beneficial effects of the present invention are：
1. combination electrode of the present invention preparation is simple and convenient to operate, manufacturing cost is low, detection speed is fast, stability and reproduction
Property is good.
2. combination electrode of the present invention is to ranging from 10~1000 μM of the Electrochemical Detection of NADH, detection limit is down to 2.7 μM
(S/N=3).
Description of the drawings
The accompanying drawings which form a part of this application are used for providing further understanding of the present application, and the application's shows
Meaning property embodiment and its explanation do not constitute the improper restriction to the application for explaining the application.
Fig. 1 is the scanning electron microscope (SEM) photograph of combination electrode prepared by embodiment 1, and A is carbon paper, and B is combination electrode；
Fig. 2 is the cyclic voltammogram that combination electrode prepared by embodiment 1 detects NADH；
Fig. 3 is the differential pulse voltammetry curve that combination electrode prepared by embodiment 1 detects NADH, and a is that NADH is a concentration of
10 μM, i is a concentration of 1000 μM of NADH.
Specific implementation mode
It is noted that described further below be all exemplary, it is intended to provide further instruction to the application.Unless another
It indicates, all technical and scientific terms used herein has usual with the application person of an ordinary skill in the technical field
The identical meanings of understanding.
It should be noted that term used herein above is merely to describe specific implementation mode, and be not intended to restricted root
According to the illustrative embodiments of the application.As used herein, unless the context clearly indicates otherwise, otherwise singulative
It is also intended to include plural form.Additionally, it should be understood that when using term "comprising" and/or " packet in the present specification
Include " when, indicate existing characteristics, step, operation, device, component and/or combination thereof.
Electrochemical deposition method described herein is in the solution containing gold ion, using the substrate of required precipitation as work
Make electrode, inertia resistant material precipitates the process of gold nano grain by being electrolysed as auxiliary electrode in substrate.
As background technology is introduced, the ineffective deficiency of existing electrode detection NADH exists in the prior art,
In order to solve technical problem as above, present applicant proposes the electrifications that a kind of carbon paper-gold nano grain combination electrode detects NADH
Method.
It is compound to provide a kind of carbon paper-gold nano grain for detecting NADH for a kind of exemplary embodiment of the application
Gold nano grain is deposited on carbon paper at least 1 using electrochemical deposition method and enclosed, you can obtained for detecting by the preparation method of electrode
The carbon paper of NADH-gold nano grain combination electrode.
Combination electrode prepared by the application, stability is high, preparation process is simple, can realize the highly sensitive inspection of NADH
It surveys.
Preferably, step is：
(1) gold chloride is evenly dispersed to obtaining homogeneous phase solution in solvent；
(2) using carbon paper as working electrode, by three-electrode system after homogeneous phase solution carries out the circle of electrochemical deposition at least 1
Obtain combination electrode.
It is further preferred that the solvent in step (1) is secondary water or phosphate buffer solution (PBS).It is herein described
Secondary water be redistilled water.
It is further preferred that in homogeneous phase solution gold chloride a concentration of 0.1~0.5mM.It is experimentally confirmed, under the concentration
The detection result of the combination electrode of preparation is best.
In order to keep gold chloride evenly dispersed in a solvent, it is further preferred that step (1) uses ultrasound in dispersion process
Processing.Still more preferably, the time of supersound process is 5~15min.
It is further preferred that the method for electrochemical deposition is cyclic voltammetric (CV) method in step (2).Working electrode is carbon
Paper, and carbon paper is fixed by electrode holder.
Still more preferably, the parameter of step (2) is：Potential range -1~1V sweeps 5~50mVs of speed-1, deposition circle
Number is 1~15 circle.
The step of combination electrode is activated before use, activation is that combination electrode is existed using cyclic voltammetry
0.5M H2SO4In solution, with 50mV s-1Sweep speed 0.2~1.5V (vs.SCE) scanning 10 circle.
The another embodiment of the application provides a kind of carbon for detecting NADH prepared by above-mentioned preparation method
Paper-gold nano grain combination electrode.
The application the third embodiment there is provided it is a kind of detection NADH electrochemical method, using above-mentioned electrode as work
Make electrode, the solution containing NADH is detected using cyclic voltammetry (CV) or differential pulse voltammetry (DPV).
Preferably, cyclic voltammetry testing conditions are：Potential range -0.2~0.8V (vs.SCE) is detected, fast 50mV is swept
s-1。
Differential pulse voltammetry testing conditions are：Detect 0~0.8V of potential range, pulse height 2.5mV, pulse width
For 100ms, step height 5mV, snap time 500ms.The electricity of cyclic voltammetry (CV) or differential pulse voltammetry (DPV)
It is 0.1M PBS to solve liquid.
In order to enable those skilled in the art can clearly understand the technical solution of the application, below with reference to tool
The technical solution of the application is described in detail in the embodiment of body.
Embodiment 1
(1) novel carbon paper-gold nano grain combination electrode (CP-AuNxE preparation).First, carbon paper is cut into fixed
Shape is thoroughly cleaned successively with absolute ethyl alcohol and secondary water after being clipped with electrode holder, nitrogen is used in combination to dry up, for use.Then, by chlorine
Auric acid is diluted to a concentration of 0.5mM with secondary water.By above-mentioned electrode directly as working electrode, platinum electrode is auxiliary electrode,
Saturated calomel electrode (SCE) is reference electrode, while being immersed in the chlorauric acid solution of 0.5mM, and electricity is carried out on the surfaces CP with CV methods
Chemical deposition is tested.Experiment parameter is arranged：Potential range -1~1V sweeps fast 50mVs-1, the deposition circle of the number of turns 7.Obtain experiment institute
Novel carbon paper-gold nano grain combination electrode is labeled as CP-AuN7E.Carbon paper (A) and carbon paper-gold nano is shown in Fig. 1
The scanning electron microscope (SEM) photograph of Particles dispersed electrode (B).Gold nanoparticle is evenly distributed in carbon paper surface as can see from Figure 1, and
The size and pattern of the gold nanoparticle of deposition can be controlled by number of deposition cycles.
(2) carbon paper-gold nano grain combination electrode (CP-AuN7E) Electrochemical Detection of the electrode to NADH.First, pH is used
The NADH solution that=7.0 0.1M phosphate buffer solutions (PBS) directly compound concentration is 1mM, then use respectively CP electrodes and
CP-AuN7E is detected NADH solution by CV methods, experiment parameter setting：- 0.2~0.8V (vs.SCE), sweeps fast 50mV
s-1.Experimental result is as shown in Fig. 2, CP-AuN7E is much more obvious to the response ratio CP electrodes of NADH.
NADH is diluted to various concentration with the phosphate buffer solution of pH=7.0, then uses CP-AuN7E is by the side DPV
Method is detected.Detection parameters are arranged：0~0.8V, pulse height 2.5mV, pulse width 100ms, step height are
5mV, snap time 500ms.Experimental result is as shown in figure 3, the combination electrode detects linear ranging from 10~1000 μ to NADH
M, detection are limited to 2.7 μM (S/N=3).
With the same CP-AuNPs electrodes to the NADH solution METHOD FOR CONTINUOUS DETERMINATION 6 times of 1mM, it is found that peak current varies less, RSD
It is 2.1%, shows that CP-AuNPs electrodes have excellent reproducibility.Meanwhile we also study under experiment condition after optimization
The reproducibility of 6 CP-AuNPs electrodes, RSD 3.2%.In addition, the electrode tested is rinsed with secondary water, nitrogen drying
After postposition stores 2 weeks at room temperature, CP-AuNPs electrodes still remain initial value to the oxidation peak current of the NADH solution of 1mM
97.6%, it was confirmed that the Novel paper base electrode has excellent stability.
Embodiment 2.
Novel carbon paper-gold nano grain combination electrode (CP-AuN7E preparation).
First, carbon paper is cut into fixed shape, uses absolute ethyl alcohol and secondary water thoroughly clear successively after being clipped with electrode holder
It washes, nitrogen is used in combination to dry up, for use.Then, gold chloride is diluted to a concentration of 0.1mM with secondary water.Above-mentioned electrode is directly made
For working electrode, platinum electrode is auxiliary electrode, and saturated calomel electrode (SCE) is reference electrode, while immersing the chlorine gold of 0.1mM
In acid solution, electrochemical deposition experiment is carried out on the surfaces CP with CV methods.Experiment parameter is arranged：Potential range -1~1V, sweeps speed
10mV·s-1, the deposition circle of the number of turns 7.It obtains testing carbon paper used-gold nano grain combination electrode (CP-AuN7E), 7 represent not
The synsedimentary number of turns.
Embodiment 3.
Novel carbon paper-gold nano grain combination electrode (CP-AuN10E preparation).
First, carbon paper is cut into fixed shape, uses absolute ethyl alcohol and secondary water thoroughly clear successively after being clipped with electrode holder
It washes, nitrogen is used in combination to dry up, for use.Then, gold chloride is diluted to a concentration of 0.5mM with PBS.By above-mentioned electrode directly as work
Make electrode, platinum electrode is auxiliary electrode, and saturated calomel electrode (SCE) is reference electrode, while the gold chloride for immersing 0.5mM is molten
In liquid, electrochemical deposition experiment is carried out on the surfaces CP with CV methods.Experiment parameter is arranged：Potential range -1~1V sweeps fast 10mV
s-1, the deposition circle of the number of turns 10.It obtains testing carbon paper used-gold nano grain combination electrode (CP-AuN10E), 10 different sink is represented
The product number of turns.
Embodiment 4.
Novel carbon paper-gold nano grain combination electrode (CP-AuN5E preparation).
First, carbon paper is cut into fixed shape, uses absolute ethyl alcohol and secondary water thoroughly clear successively after being clipped with electrode holder
It washes, nitrogen is used in combination to dry up, for use.Then, gold chloride is diluted to a concentration of 2mM with secondary water.By above-mentioned electrode directly as
Working electrode, platinum electrode are auxiliary electrode, and saturated calomel electrode (SCE) is reference electrode, while the gold chloride for immersing 2mM is molten
In liquid, electrochemical deposition experiment is carried out on the surfaces CP with CV methods.Experiment parameter is arranged：Potential range -1~1V sweeps fast 50mV
s-1, the deposition circle of the number of turns 5.It obtains testing novel carbon paper used-gold nano grain combination electrode (CP-AuN5E), 5 difference is represented
Deposit the number of turns.
The foregoing is merely the preferred embodiments of the application, are not intended to limit this application, for the skill of this field
For art personnel, the application can have various modifications and variations.Within the spirit and principles of this application, any made by repair
Change, equivalent replacement, improvement etc., should be included within the protection domain of the application.
Claims (10)
1. a kind of preparation method for detecting carbon paper-gold nano grain combination electrode of NADH, characterized in that use electrochemistry
Gold nano grain is deposited on carbon paper at least 1 and enclosed by sedimentation, you can is obtained compound for detecting carbon paper-gold nano grain of NADH
Electrode.
2. preparation method as described in claim 1, characterized in that step is：
(1) gold chloride is evenly dispersed to obtaining homogeneous phase solution in solvent；
(2) using carbon paper as working electrode, in homogeneous phase solution obtain after electrochemical deposition at least 1 encloses by three-electrode system
Combination electrode.
3. preparation method as claimed in claim 2, characterized in that the solvent in step (1) is secondary water or phosphate-buffered
Solution.
4. preparation method as claimed in claim 2, characterized in that a concentration of 0.1~0.5mM of gold chloride in homogeneous phase solution.
5. preparation method as claimed in claim 2, characterized in that step (1) is in dispersion process using supersound process.
6. preparation method as claimed in claim 2, characterized in that the method for electrochemical deposition is cyclic voltammetric in step (2)
Method.
7. preparation method as claimed in claim 6, characterized in that the parameter of step (2) is：Potential range -1~1V sweeps speed 5
~50mVs-1, the deposition number of turns is 1~15 circle.
8. carbon paper-gold nano grain for detecting NADH prepared by a kind of any preparation method of claim 1~7 is multiple
Composite electrode.
9. a kind of electrochemical method of detection NADH, characterized in that using electrode according to any one of claims 8 as working electrode, use
Cyclic voltammetry or differential pulse voltammetry are detected the solution containing NADH.
10. electrochemical method as claimed in claim 9, characterized in that cyclic voltammetry testing conditions are：Detect current potential model
- 0.2~0.8V is enclosed, fast 50mVs is swept-1；
Differential pulse voltammetry testing conditions are：0~0.8V of potential range, pulse height 2.5mV are detected, pulse width is
100ms, step height 5mV, snap time 500ms.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111830097A (en) * | 2019-10-22 | 2020-10-27 | 中国农业科学院北京畜牧兽医研究所 | Electrochemical detection method of egg pentamethyl tetrahydrofolic acid |
| CN112083050A (en) * | 2020-09-17 | 2020-12-15 | 南京工业大学 | Preparation method of NADH and ethanol biosensor chip |
-
2018
- 2018-06-04 CN CN201810565299.3A patent/CN108614022A/en active Pending
Non-Patent Citations (4)
| Title |
|---|
| JUN HO SHIM ET AL: "The effect of electrochemical pretreatment on the catalytic activity of carbon-supported gold nanoparticles for NADH oxidation", 《MATERIALSLETTERS》 * |
| LIN TANG ET AL.: "Highly sensitive sensor for detection of NADH based on catalytic growth of Au nanoparticles on glassy carbon electrode", 《ANAL BIOANAL CHEM》 * |
| YUE WAN ET AL.: "Au nanoparticle modified carbon paper electrode for an electrocatalytic oxidation nitrite sensor", 《NEW J. CHEM.》 * |
| 李靖 等: "NADH在纳米金/过氧化聚吡咯复合材料修饰电极上的电催化氧化", 《上海第二工业大学学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111830097A (en) * | 2019-10-22 | 2020-10-27 | 中国农业科学院北京畜牧兽医研究所 | Electrochemical detection method of egg pentamethyl tetrahydrofolic acid |
| CN111830097B (en) * | 2019-10-22 | 2023-05-30 | 播恩集团股份有限公司 | Electrochemical detection method of egg pentamethyltetrahydrofolic acid |
| CN112083050A (en) * | 2020-09-17 | 2020-12-15 | 南京工业大学 | Preparation method of NADH and ethanol biosensor chip |
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