CN109406600A - It is a kind of can be in the production method of body real-time detection hydrogen peroxide microelectrode - Google Patents
It is a kind of can be in the production method of body real-time detection hydrogen peroxide microelectrode Download PDFInfo
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- CN109406600A CN109406600A CN201811093606.9A CN201811093606A CN109406600A CN 109406600 A CN109406600 A CN 109406600A CN 201811093606 A CN201811093606 A CN 201811093606A CN 109406600 A CN109406600 A CN 109406600A
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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
- 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
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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
- G01N27/308—Electrodes, e.g. test electrodes; Half-cells at least partially made of carbon
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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 discloses one kind can be in the production method of body real-time detection hydrogen peroxide microelectrode, the following steps are included: 1. modifying single-walled carbon nanotube (SWCNTs) on carbon fibre ultramicro-electrode (CFUMEs), specific steps are as follows: carbon fibre ultramicro-electrode is immersed in SWCNTs suspension, and it is placed under infrared lamp and toasts, it is added dropwise again after suspension drying, altogether three times；2. modifying horseradish peroxidase-gold nano grain compound (HRP-AuNPs) on the carbon fibre ultramicro-electrode of modified SWCNTs, specific steps are as follows: the carbon fibre ultramicro-electrode of 1. modified single-walled carbon nanotube (SWCNTs) that step obtains is immersed in HRP-AuNPs solution, modifies 12 h under the conditions of 4 DEG C.It is prepared by the present invention to reduce the wound to organism in body real-time detection hydrogen peroxide microelectrode, sensitivity and the signal-to-noise ratio of electrode are improved, real-time, the dynamic monitoring of cell or tissue microenvironment are suitble to.
Description
Technical field
The invention belongs to electrochemical analysis, electrochemical sensor technology field, and in particular to one kind can be in body real-time detection
The production method of hydrogen peroxide microelectrode.
Background technique
Hydrogen peroxide is a kind of important small molecule present in biosystem, and important work is played in many physiology courses
With.Rapidly and sensitively detecting hydrogen peroxide is the major issue that scientific circles pay close attention to for a long time.It at present include chemiluminescence
Numerous methods including method, fluorescence method and spectrophotometry and electrochemical method etc. are for detecting hydrogen peroxide, wherein electrification
Method relies on its high sensitivity, is not required to detection and analysis that the advantages such as chemical probe are widely used in hydrogen peroxide.
Enzyme biologic sensor in electrochemical method was obtained in recent years due to advantages such as sensitivity, the selectivity of enzyme itself
Greatly development.Simultaneously with the vitro detection of bioactive substance and the maturation of analytical technology, it is used for biological active matter
Matter has also attracted the concern of more and more researchers in physical examination survey.Microelectrode relies on its high sensitivity, high-resolution, high conduction
The excellent electrochemical properties such as rate analyze in the electro physiology of life science, biological cell, in terms of send out
Irreplaceable role has been waved, has adequately been paid attention to and is widely applied.Conventional microelectrode it is one-dimensional having a size of micron order,
Its biggish size is not suitable for surveying in physical examination for bioactive substance.In addition, the preparation process of microelectrode directly affects electrification
Sensitivity, the resolution ratio, repeatability, accuracy of detection are learned, while preparation process is complicated, the factors restriction microelectrode such as at high cost exists
The development of bioactive substance detection field.
In summary factor, small, high sensitivity that the invention proposes a kind of sizes, production carbon fiber simple, at low cost
The preparation method of ultramicroelectrode.I.e. using it is one-dimensional having a size of micron-sized carbon fiber as electrode, good electron transmission energy
The electrochemical properties such as power, fabulous biocompatibility, high s/n ratio, high current density be both able to satisfy the needs of electrode material；This
Its outer minimum size can reduce the wound to organism again, improve sensitivity and the signal-to-noise ratio of electrode, be suitble to cell or group
Knit real-time, the dynamic monitoring of microenvironment.
Summary of the invention
Small, high sensitivity that the object of the present invention is to provide a kind of sizes, production is simple, at low cost can be in body real-time detection
The production method of hydrogen peroxide microelectrode.
To achieve the above object, the technical solution adopted by the present invention is that, one kind can be in the micro- electricity of body real-time detection hydrogen peroxide
The production method of pole, comprising the following steps:
1. modifying single-walled carbon nanotube (SWCNTs) on carbon fibre ultramicro-electrode (CFUMEs), specific steps are as follows: by carbon fiber
Ultramicroelectrode is immersed in SWCNTs suspension, and is placed under infrared lamp and is toasted, and is added dropwise again after suspension drying, altogether three times；
2. modifying horseradish peroxidase-gold nano on the carbon fibre ultramicro-electrode of modified single-walled carbon nanotube (SWCNTs)
Particle composites (HRP-AuNPs), specific steps are as follows: the carbon for the modified single-walled carbon nanotube (SWCNTs) for 1. obtaining step
Fiber ultramicroelectrode is immersed in HRP-AuNPs solution, modifies 12 h under the conditions of 4 DEG C.
Preferably, the carbon fibre ultramicro-electrode includes carbon fiber, contact conductor and glass capillary, carbon fiber and electrode
Lead is connected by silver conductive adhesive, and one end of glass capillary is drawn into the tip (capillary glass for being 0.9 ~ 1.1 mm by internal diameter
Pipe stretched tip under alcolhol burner flame), the carbon fiber connecting with contact conductor penetrates the tip of glass capillary, and carbon fiber
Dimension exposes glass capillary tip (the outer carbon fiber length of pipe is 2 ~ 3 mm), and contact conductor exposes the other end of glass capillary,
It is sealed with resin at the end.
Preferably, the contact conductor is copper wire, and brass wire diameter is 400 ~ 600 μm.
Preferably, the length of the carbon fiber is 2 ~ 2.5 cm, and diameter is 7 μm.
Preferably, step 1. in SWCNTs suspension the preparation method is as follows: SWCNTs is divided by 0.3 mg:1mL of solid-to-liquid ratio
It dissipates in 1 mg/mL cetyl trimethylammonium bromide (CTAB) aqueous solution, 1 ~ 1.5 h of ultrasound.
Preferably, step 2. in HRP-AuNPs solution the preparation method is as follows: taking 50 mg HRP in 50 mL round-bottomed flasks
In, 5 mL deionized waters are added, then 5 mL 3mM HAuCl are added into flask4 Solution, then the NaOH solution with 1 mol/L
Adjusting pH is 12, then reacts 10 h under the conditions of temperature is 37 DEG C.
The beneficial effect comprise that: the present invention is used as electrode having a size of micron-sized carbon fiber using one-dimensional, logical
Modification single-walled carbon nanotube (SWCNTs) and horseradish peroxidase-gold nano grain compound (HRP-AuNPs) are crossed, is prepared
Microelectrode have the electrochemistry such as good electron transmission ability, fabulous biocompatibility, high s/n ratio, high current density special
Property, it can be very good the needs for meeting electrode material；Furthermore its minimum size can reduce the wound to organism again, improve
The sensitivity of electrode and signal-to-noise ratio are suitble to real-time, the dynamic monitoring of cell or tissue microenvironment.
Detailed description of the invention
Fig. 1 can be in the structural schematic diagram of body real-time detection hydrogen peroxide microelectrode for of the invention；In figure: 1 is carbon fiber,
2 be silver conductive adhesive, and 3 be copper wire, and 4 be glass capillary, and 5 be resin；
Fig. 2 is in PBS(pH=7.0 of 0.01 M) in, shown with cyclic voltammetry electrochemical activation carbon fibre ultramicro-electrode process
It is intended to；Platinum electrode is to electrode, and Ag/AgCl(is saturated KCl) electrode is reference electrode, scanning current potential is -0.2 ~+1.2 V, is swept
Retouching rate is 100 mVs-1；
Fig. 3 is HRP-AuNPs/SWCNTs/CFUME(a), HRP/SWCNTs/CFUME(b), SWCNTs/CFUME(c), HRP-
AuNPs/CFUME(d), HRP/CFUME(e) and CFUME(f) in PBS(pH=7.0 of 0.01 M, anaerobic) in, in sweep speed
For 100 mVs-1When cyclic voltammetry curve；Platinum electrode is to electrode, and Ag/AgCl(is saturated KCl) electrode is reference electrode, scanning
Current potential is -0.6 ~+0.2 V；
Fig. 4 is HRP-AuNPs/SWCNTs/CFUME in PBS(pH=7.0 of 0.01 M, anaerobic) in, it is 10 in sweep speed,
50,100,150,200,250,300,350,400,450 and 500mVs-1When cyclic voltammetry curve；Platinum electrode be to electrode,
Ag/AgCl(be saturated KCl) electrode be reference electrode, scanning current potential be -0.6 ~+0.2 V；Illustration is anode and cathode peak electricity
Flow the figure to sweep speed；
Fig. 5 is HRP-AuNPs/SWCNTs/CFUME(A), HRP/SWCNTs/CFUME(B), SWCNTs/CFUME(C), HRP-
AuNPs/CFUME(D), HRP/CFUME(E) and CFUME(F) in PBS(pH=7.0 of 0.01 M, anaerobic) in, it is not depositing
In H2O2(a), there are 0.01 mM H2O2(b) and there are 0.03 mM H2O2(c) cyclic voltammetry curve when；Platinum electrode is
To electrode, Ag/AgCl(is saturated KCl) electrode is reference electrode, scanning current potential is -0.6 ~+0.2 V, sweep speed 100
mVs-1；
Fig. 6 is HRP-AuNPs/SWCNTs/CFUME(a), HRP/SWCNTs/CFUME(b), SWCNTs/CFUME(c),
HRP-AuNPs/ CFUME(d), HRP/CFUME(e) and CFUME(f) in PBS(pH=7.0 of 0.01 M, anaerobic) in, it is right
0.01 mM H2O2Ampere response；Application voltage is -0.1V(vs.Ag/AgCl sat. KCl)；
Fig. 7 be HRP-AuNPs/SWCNTs/CFUME in PBS(pH=7.0 of 0.01M, anaerobic) in 0.01 mM H2O2,
0.09 mM glutathione (GSH) (Dextrose Monohydrate (GLU), ascorbic acid (AA), Dopamine hydrochloride (DA) or glycine
And 0.01 mM H (GLy))2O2Ampere response；Application voltage is -0.1V(vs. Ag/AgCl sat. KCl)；
Fig. 8 be in PBS(pH=7.0 of 0.01M, anaerobic) in H2O2The relation curve of concentration and response current, apply voltage be-
0.1V(vs.Ag/AgCl sat. KCl)；
Fig. 9 is in PBS(pH=7.0 with 0.01M, anaerobic) in ten times of SD rat blood serum of dilution, H2O2Reduction current and its
The relation curve of concentration, application voltage are -0.1V(vs. Ag/AgCl sat. KCl)；
Figure 10 be HRP-AuNPs/SWCNTs/CFUME to from be added 5.6 μM of N- formyl-L- methionyl-L- leucyl-L-
Mankind's menses of buffer (b line) of the mankind of phenylalanine (fMLP) (a line) through hemocytoblast, addition without fMLP are dry thin
The ampere response of the plain buffer (c line) of 5.6 μM of fMLP of born of the same parents and addition.
Specific embodiment
The present invention is further explained in the light of specific embodiments, and but the scope of the present invention is not limited thereto.
Embodiment 1
1. the preparation of HRP-AuNPs solution:
(1) 10 h of chloroazotic acid bulb glass instrument is used, cleaned, dried with deionized water later；
(2) oil bath pan is opened, 1 h is preheated, final temperature is 37 DEG C；
(3) it takes 50 mg HRP in 50 mL round-bottomed flasks, 5 mL deionized waters is added, then 5 mL 3mM are added into flask
HAuCl4 0.45 mL 1M NaOH solution (adjusting pH value of solution is 12) is added in solution into flask after ten minutes, and flask is placed in
In oil bath pan, solution reacts 10 h under stirring.
2. the production and modification of carbon fibre ultramicro-electrode:
(1) copper wire is taken, it is with sand paper that one end polishing is bright, it is cleaned by ultrasonic respectively in deionized water, acetone, deionized water
Three minutes, taking-up was dried；
(2) carbon fiber is fixed on one end that copper wire is polished with silver conductive adhesive；
(3) by glass capillary that internal diameter is 1.0 mm under alcolhol burner flame stretched tip, carbon fiber-copper wire is from capillary
It is inserted into and passes through tip, and one not drawn with resin (AB glue) fixed copper wire and glass capillary in one end that glass tube is not drawn
End；
(4) capillary tip melts sealing under alcolhol burner flame, and carbon fiber is fixed in capillary, and it is outer to cut pipe with scissors
Carbon fiber length is 2.5 mm, obtains carbon fibre ultramicro-electrode, as shown in Figure 1；
(5) with deionized water, acetone, deionized water be cleaned by ultrasonic carbon fibre ultramicro-electrode, three minutes every time；
(6) with cyclic voltammetry in 0.01 PBS(pH=7.0 M) in activated carbon fiber ultramicroelectrode, sweep speed 100
mVs-1, scanning current potential is -0.2 ~+1.2 V, 40 circle of scanning；Ag/AgCl(is saturated KCl) electrode is reference electrode, platinum electrode is
Auxiliary electrode, as shown in Figure 2；
(7) activated carbon fibre ultramicro-electrode is taken out and is cleaned with deionized water, being immersed in SWCNTs suspension, (SWCNTs is outstanding
Liquid the preparation method comprises the following steps: disperse SWCNTs in 1mg/mL CTAB aqueous solution by 0.3 mg:1mL of solid-to-liquid ratio, ultrasound 1.5 is h)
In, and be placed under infrared lamp and toast, it is added dropwise again after suspension drying, altogether three times；
(8) carbon fibre ultramicro-electrode of modified single-walled carbon nanotube (SWCNTs) in (7) is immersed in HRP-AuNPs solution
In, 12 h are modified under the conditions of 4 DEG C.
Embodiment 2
1. Direct Electrochemistry behavior of the HRP-AuNPs on HRP-AuNPs/SWCNTs/CFUMEs
In PBS(pH=7.0 of 0.01 M, anaerobic) in, the electrification of fixed HRP-AuNPs is characterized using cyclic voltammetry
Scholarship and moral conduct is.Fig. 3 shows HRP-AuNPs/SWCNTs/CFUME(a respectively), HRP/SWCNTs/CFUME(b), SWCNTs/CFUME
(c), HRP-AuNPs/CFUME(d), HRP/CFUME(e) and naked carbon fibre ultramicro-electrode CFUME(f) cyclic voltammetric it is bent
Line.In Fig. 3 d, e, f, in the potential range of -0.6 ~+0.2V, HRP-AuNPs/CFUME, HRP/CFUME and naked
CFUME does not detect redox peaks.Apparent redox peaks are not detected in Fig. 3 c, but since single wall carbon is received
The modification of mitron, the electric current detected significantly increase, and can be attributed to the surface area of the increase of single-walled carbon nanotube and good
Electric conductivity, which form the effective roads of the direct electron transfer between many horseradish peroxidases and carbon fibre ultramicro-electrode
Diameter.Detecting apparent redox peaks in Fig. 3 a, oxidation spike potential is -0.096 V, reduction spike potential is -
0.215 V, it is peak-to-peak to be divided into 119 mV, show a kind of Quasi-reversible process.This is by horseradish peroxidase to hydrogen reduction peak
What prothetic group Fe (III)/Fe (II) redox couple generated, the successful fixation of HRP-AuNPs is shown, this is attributed to the fact that
The modification of SWCNTs, and Fig. 3 b does not detect apparent redox peaks, shows horseradish peroxidase and Jenner
The combination of rice grain significantly enhances the electron transfer efficiency of enzyme and electrode surface.To further characterize HRP-AuNPs/
The electrochemical behavior of SWCNTs/CFUMEs, in PBS(pH=7.0 of 0.01 M, anaerobic) in, different cyclic voltammetry scan speed
Rate is applied, and current-responsive is recorded.Fig. 4 shows HRP-AuNPs/SWCNTs/CFUME in 10 ~ 500 mVs-1Scanning
Cyclic voltammetry curve under rate.As shown in Fig. 4 illustration, oxidation peak and reduction peak current are all as the increase of sweep speed is in line
Property increase, related coefficient (R2) it is 0.996, showing a kind of typical granule surface contral will definitely inverse electrochemical process.These knots
Fruit meets can this special adsorption layer equation (ip=n2F2VA Γ */RT), wherein ipIt is peak current, n is electron transmission quantity, and F is
Faraday constant, R are gas constants, and T is absolute temperature, and v is sweep speed, and A is the effective surface area of electrode, Γ *
It is the surface concentration of electroactive material.Electroactive material according to the equation, on HRP-AuNPs/SWCNTs/CFUMEs
The surface coverage value (Γ *) of HRP-AuNPs molecule is estimated as 5.15 × 10-9 mol cm-2, than unimolecule coverage rate (1.89
×10-11 mol cm-2) high two orders of magnitude.This is the result shows that SWCNTs is fixed on CFUMEs with very HRP-AuNPs
Good facilitation effect, and effectively mediate the electron transmission from HRP-AuNPs to CFUMEs.
2. H on HRP-AuNPs/SWCNTs/CFUMEs2O2Electro-catalysis reduction
HRP-AuNPs/SWCNTs/CFUMEs is to H2O2Electro-catalysis reducing property characterized first by cyclic voltammetry.?
There is no (a), there are 0.01 mM(b) and 0.03 mM(c) H2O2In the case where HRP-AuNPs/SWCNTs/CFUME(A), HRP/
SWCNTs/CFUME(B), SWCNTs/CFUME(C), HRP-AuNPs/CFUME(D), HRP/CFUME(E) and CFUME(F) 8
PBS(pH=7.0 of 0.01 M of mL, anaerobic) in current-responsive be recorded such as Fig. 5.H is being added2O2Later, HRP-AuNPs/
SWCNTs/CFUME(A reduction current) significantly increases, and HRP/SWCNTs/CFUME(B), SWCNTs/CFUME(C), HRP-
AuNPs/CFUME(D), HRP/CFUME(E) and reduction current CFUME(F) there is no variation, show these types of electrode
To H2O2It there is no electro-catalysis reducing power.These the result shows that: HRP-AuNPs/SWCNTs/CFUMEs is to H2O2Have good
Good biological electro catalysis ability.Further to study H2O2Electro-catalysis reduction on HRP-AuNPs/SWCNTs/CFUMEs,
HRP-AuNPs/SWCNTs/CFUME(a), HRP/SWCNTs/CFUME(b), SWCNTs/CFUME(c), HRP-AuNPs/CFUME
(d), HRP/CFUME(e) and CFUME(f) in PBS(pH=7.0 of 0.01M, anaerobic) in, applying current potential -0.1V
Under conditions of (vs. Ag/AgCl sat. KCl), to the H of 0.01 mM of addition2O2Response it is as shown in Figure 6.Lower electricity
Position can effectively reduce possible interference, therefore to H2O2There is relatively high selectivity.In Fig. 6, HRP-AuNPs/
SWCNTs/CFUME(a) to H2O2Current-responsive ratio HRP/SWCNTs/CFUME(b) more than 30 times big, SWCNTs/CFUME(c),
HRP-AuNPs/CFUME(d), HRP/CFUME(e) and CFUME(f) do not respond to.These results indicate that in H2O2 Ampere
In detection, HRP-AuNPs/SWCNTs/CFUMEs is compared to other five kinds of electrodes with overwhelming advantage.
3. detection of the HRP-AuNPs/SWCNTs/CFUMEs to different activities substance anti-interference ability
In PBS(pH=7.0 of 0.01M, anaerobic) in, in the item for applying current potential -0.1V (vs. Ag/AgCl sat. KCl)
Under part, to the H of 0.01 mM of addition2O2It is as shown in Figure 7 with the current-responsive of 0.09 mM chaff interferent.The results show that 9 times
H2O2The glutathione (GSH) of concentration, Dextrose Monohydrate (GLU), ascorbic acid (AA), Dopamine hydrochloride (DA) and glycine
(GLy) without any disturbance response, the H of 0.01 mM2O2Response be not interfered the influence of object yet.These results card
Real, HRP-AuNPs/SWCNTs/CFUMEs has good anti-interference ability, to H2O2With very high selectivity.
Embodiment 3
1. HRP-AuNPs/SWCNTs/CFUMEs is to H2O2Vitro detection and its stability
In PBS(pH=7.0 of 0.01M, anaerobic) in, in the item for applying current potential -0.1V (vs. Ag/AgCl sat. KCl)
Under part, HRP-AuNPs/SWCNTs/CFUME is detected to H by current time method2O2Response, record reduction current, such as Fig. 8 institute
Show.Its range of linearity is 2.4 μM ~ 34 μM (R2=0.991), detection be limited to 443 nM(S/N=3), should the result shows that
HRP-AuNPs/SWCNTs/CFUMEs is to H2O2With extremely strong detectability.One in the environment of modified electrode is placed on 4 DEG C
Week, to H2O2Response almost without reduction, show that HRP-AuNPs/SWCNTs/CFUMEs has long time-histories stability.
2. HRP-AuNPs/SWCNTs/CFUMEs is in SD rat blood serum to H2O2Detection
In PBS(pH=7.0 with 0.01M, anaerobic) ten times of dilution SD rat blood serum in, applying current potential -0.1V (vs.
Ag/AgCl sat. KCl) under conditions of, HRP-AuNPs/SWCNTs/CFUME is detected to H by current time method2O2Sound
It answers, records reduction current, as shown in Figure 9.Its range of linearity is 10 μM ~ 268.8 μM (R2=0.997), detection is limited to 2.5
μM (S/N=3), should the result shows that, the detection limit of HRP-AuNPs/SWCNTs/CFUMEs decreases in serum, but to H2O2
Still there is very strong detectability, provides basis in physical examination survey for electrode.
3. HRP-AuNPs/SWCNTs/CFUMEs discharges H through hemocytoblast to the mankind2O2Detection
It can produce H under the stimulation of fMLP2O2The mankind be chosen as mode cell through hemocytoblast to characterize HRP-AuNPs/
The Electrochemical Detection ability of SWCNTs/CFUMEs.Figure 10 is shown in 1 PBS(pH=7.0 mL 0.01M) in, in application-
0.1 V(vs. Ag/AgCl sat. KCl) voltage when current-time curvel.After baseline stability, 5.6 are added into solution
μM fMLP(a line, Figure 10).As control, under kindred circumstances, the buffer solution (c line, Figure 10) of cell is not added same dense
The fMLP of degree and in cell plus two systems of buffer solution (b line, Figure 10) of blank are also detected, it is tested without curent change
It measures.In the case where there is stem cell, electric current is obviously increased after fMLP is added.These are the result shows that HRP-AuNPs/SWCNTs/
CFUMEs can be used for detecting the H of cell release2O2。
Claims (6)
1. one kind can be in the production method of body real-time detection hydrogen peroxide microelectrode, it is characterised in that the following steps are included:
1. modifying single-walled carbon nanotube SWCNTs, specific steps are as follows: soak carbon fibre ultramicro-electrode on carbon fibre ultramicro-electrode
Bubble is placed under infrared lamp and toasts in SWCNTs suspension, is added dropwise again after suspension drying, altogether three times；
2. modifying HRP-AuNPs, specific steps on the carbon fibre ultramicro-electrode of modified single-walled carbon nanotube SWCNTs are as follows: will
The carbon fibre ultramicro-electrode of 1. modified SWCNTs that step obtains is immersed in HRP-AuNPs solution, is repaired under the conditions of 4 DEG C
Adorn 12 h.
2. as described in claim 1 can be in the production method of body real-time detection hydrogen peroxide microelectrode, which is characterized in that the carbon
Fiber ultramicroelectrode includes carbon fiber, contact conductor and glass capillary, and carbon fiber is connect with contact conductor by silver conductive adhesive,
One end of glass capillary is drawn into tip, and the carbon fiber connecting with contact conductor penetrates the tip of glass capillary, and carbon fiber
Dimension exposes glass capillary tip, and contact conductor exposes the other end of glass capillary, which is sealed with resin.
3. as claimed in claim 2 can be in the production method of body real-time detection hydrogen peroxide microelectrode, which is characterized in that the electricity
Pole lead is copper wire, and brass wire diameter is 400 ~ 600 μm.
4. as claimed in claim 2 can be in the production method of body real-time detection hydrogen peroxide microelectrode, which is characterized in that the carbon
The length of fiber is 2 ~ 2.5 cm, and diameter is 7 μm.
5. as described in claim 1 can be in the production method of body real-time detection hydrogen peroxide microelectrode, it is characterised in that step is 1.
Middle SWCNTs suspension the preparation method is as follows: dispersing 1 mg/mL CTAB aqueous solution by 0.3 mg:1mL of solid-to-liquid ratio for SWCNTs
In, 1 ~ 1.5 h of ultrasound.
6. as described in claim 1 can be in the production method of body real-time detection hydrogen peroxide microelectrode, it is characterised in that step is 2.
Middle HRP-AuNPs solution the preparation method is as follows: take 50 mg HRP in 50 mL round-bottomed flasks, 5 mL deionized waters are added,
5 mL 3mM HAuCl are added into flask again4 Solution, then adjusting pH with the NaOH solution of 1 mol/L is 12, then in temperature
Degree reacts 10 h under the conditions of being 37 DEG C.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112748163A (en) * | 2019-10-30 | 2021-05-04 | 中国科学院化学研究所 | Electrochemical sensor synchronously recording with electrophysiology |
| CN113008957A (en) * | 2021-01-20 | 2021-06-22 | 新乡医学院 | Method for manufacturing double microelectrodes capable of detecting hydrogen peroxide and nitric oxide in vivo synchronously |
| CN114486860A (en) * | 2022-01-04 | 2022-05-13 | 东南大学 | Hydrogen peroxide real-time in-situ quantitative analysis method based on bipolar nano electrode array |
| CN115248237A (en) * | 2021-04-28 | 2022-10-28 | 河南广播电视大学 | Copper-based MOF material modified microelectrode and application thereof in superoxide anion detection |
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| CN113008957A (en) * | 2021-01-20 | 2021-06-22 | 新乡医学院 | Method for manufacturing double microelectrodes capable of detecting hydrogen peroxide and nitric oxide in vivo synchronously |
| CN115248237A (en) * | 2021-04-28 | 2022-10-28 | 河南广播电视大学 | Copper-based MOF material modified microelectrode and application thereof in superoxide anion detection |
| CN115248237B (en) * | 2021-04-28 | 2024-04-26 | 河南广播电视大学 | Copper-based MOF material modified microelectrode and application thereof in superoxide anion detection |
| CN114486860A (en) * | 2022-01-04 | 2022-05-13 | 东南大学 | Hydrogen peroxide real-time in-situ quantitative analysis method based on bipolar nano electrode array |
| CN114486860B (en) * | 2022-01-04 | 2024-01-30 | 东南大学 | Hydrogen peroxide real-time in-situ quantitative analysis method based on bipolar nano electrode array |
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