CN107389755A - Electrochemical sensor for detecting mercury and its preparation method and application - Google Patents
Electrochemical sensor for detecting mercury and its preparation method and application Download PDFInfo
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Abstract
The invention provides a kind of electrochemical sensor for being used to detect mercury and its preparation method and application, electrochemical sensor includes glass-carbon electrode, probe P2 and the probe P3 for being fixed on sulfydryl modification on nano Au particle；The test side surface modification of glass-carbon electrode has carbon doping class graphene phase carbon nitride, deposition has nano Au particle on carbon doping class graphene phase carbon nitride, the probe P1 of sulfydryl modification is connected on the nano Au particle, probe P2 parts deoxyribonucleotide can form duplex structure with the probe P1 parts deoxyribonucleotide complementary pairing of sulfydryl modification, and unpaired deoxyribonucleotide can form duplex structure with being fixed on the probe P3 of sulfydryl modification on nano Au particle in probe P1 and probe P2.Its preparation method includes the steps such as modification carbon doping class graphene phase carbon nitride, modified nano gold, modification probe P1.The ability of the electrochemical sensor preventing from heavy metal ion interference of the present invention is strong, can be applied to detect mercury ion.
Description
Technical field
The present invention relates to gene technology field, more particularly to a kind of electrochemical sensor and its preparation side for being used to detect mercury
Method and application.
Background technology
At present, the method for the pollutant in determination of the environment mainly have chromatography, ultraviolet spectrometry, Synchronous fluorimetric method,
AAS, derivative spectrophotometry, flow injection analysis etc..All there is complex pretreatment in these methods, time-consuming, sample base
The defects of bulk effect is big, analytical cycle is long, higher requirement is respectively provided with to the operation level of instrument and staff, it is difficult in
Popularization and application in small business.Such as：During using spectrophotometry pollutant, because the requirement to substrate turbidity and light are done
The influence of material is disturbed, limits its accuracy and use range；And the detection of liquid and gas chromatographies is used, needed before detection pair
Sample is separated, and separation process usually requires to pre-process, and operating procedure is comparatively laborious and time-consuming, and detecting instrument is relatively expensive,
And not Portable belt, it is impossible to detected in real time.
Electrochemica biological sensor is to be based on biological organic composition (such as enzyme, antibody, nucleic acid, cell, microorganism), is treated
Examine material and carry out single-minded identification, caused signal is changed into electric signal, optical signal, and then quantitative detection by signal transduction device
Go out a new technology of test substance.Detect the heavy metal in environment, pathogenic microorganism with electrochemica biological sensor, have
Evil organic matter has the characteristics of high specificity, detection sensitivity are high, detection efficiency is high, cost is cheap, therefore becomes environment guarantor
A study hotspot in nurse's work.
At present, scientific research personnel modifies electrochemical sensor to improve electrochemical biosensor by using various new materials
Stability, repeatability and the reliability of structure of device.The key for making electrochemical DNA biosensor is how efficiently on gold
Fixing DNA probe, and it is the premise that sensor can detect to keep its activity.Usual DNA probe is in the side that gold surface is fixed
Method has by modifying DNA probe and the directly fixed two methods of not modifying DNA probe.These methods exist it is fixed it is insecure,
The defects of influenceing DNA activity using various affine materials in fixation and be easily accessible environmentally harmful material.
The content of the invention
The technical problem to be solved in the present invention is overcome the deficiencies in the prior art, there is provided one kind making is simple, stability is good,
Sensitivity and accuracy of detection are high, the strong electrification for being used to detect mercury of the ability that the common heavy metal ion of other in environment resistant is disturbed
Sensor is learned, correspondingly provides a kind of preparation method of electrochemica biological sensor, a kind of technique is simple, it is fast to make will pass through
The preparation method of speed makes fixed DNA probe have the holding of more preferable stability and high activity；On this basis, one is also provided
The application of kind foregoing electrochemical sensor, can be to simplify operation, quick response, high measurement accuracy and compared with common-path interference by force etc.
Feature realizes the efficient detection to lead in water body.
In order to solve the above technical problems, the present invention provide it is a kind of be used to detecting the electrochemical sensor of mercury, including one three
It is used as the glass-carbon electrode of working electrode in electrode system, the test side surface modification of the glass-carbon electrode has carbon doping class graphene
Phase carbon nitride, deposition has nano Au particle on the carbon doping class graphene phase carbon nitride, and the probe P1 of sulfydryl modification is connected to
On the nano Au particle, the electrochemical sensor also includes probe P2 and is fixed on the spy of sulfydryl modification on nano Au particle
Pin P3, the probe P2 parts deoxyribonucleotide can lead to the probe P1 parts deoxyribonucleotide of the sulfydryl modification
Cross thymidine-mercury-thymidine complementary pairing and form duplex structure, unpaired deoxidation core in the probe P1 and probe P2
Ribotide can form duplex structure with being fixed on the probe P3 of sulfydryl modification on nano Au particle by base pair complementarity.
Above-mentioned electrochemical sensor, it is preferred that the probe P1 is the nucleotides sequence shown in SEQ ID NO.1
Row；The probe P2 is the nucleotide sequence shown in SEQ ID NO.2；The probe P3 is with SEQ ID NO.3 institute
The nucleotide sequence shown.
Above-mentioned electrochemical sensor, it is preferred that be also embedded in the spy including signal designation agent, the signal designation agent
The duplex structure that duplex structure, probe P1 and the probe P3 that pin P1 and probe P2 is formed are formed, and probe P2 and probe P3 shapes
Into duplex structure in.
Above-mentioned electrochemical sensor, it is preferred that the signal designation agent is methylene blue, and the methylene blue concentration is
0.1~0.5mM.
The technical concept total as one, present invention also offers a kind of preparation of preceding claim electrochemical sensor
Method, comprise the following steps：
S1, carbon doping class graphene phase carbon nitride is configured to suspension, the detection end surfaces for being added drop-wise to glass-carbon electrode obtain
To the glass-carbon electrode of carbon doping class graphene phase carbon nitride modification；
S2, nano Au particle is electrodeposited in carbon doping class graphene phase carbon nitride modification glass-carbon electrode detection
End surfaces obtain the glass-carbon electrode of nanogold/carbon doping class graphene phase carbon nitride modification；
S3, the probe P1 by the glass-carbon electrode insertion sulfydryl modification of nanogold/carbon doping class graphene phase carbon nitride modification
In, the probe P1 of the sulfydryl modification is by Electrostatic Absorption on nano Au particle；It is inserted into mercaptoethanol solution, makes mercapto
Base ethanol closes nanogold not to be adsorbed, obtains probe P1/ nanogold/carbon doping class graphene phase carbon nitride modification glass carbon electricity
Pole.
Above-mentioned preparation method, it is preferred that also include matching somebody with somebody manufacturing probe P2 solution and being fixed on sulfydryl on nano Au particle repairing
The probe P3 solution of decorations；Concretely comprising the following steps for the probe P3 solution of sulfydryl modification on nano Au particle is fixed on described in preparation：Will
Ultra-pure water, chlorauric acid solution and sodium citrate solution are mixed, and dark place reaction is placed in after adding ice sodium borohydride solution while stirring
Obtain nanogold particle solution；Probe P3 and the nanogold particle solution are mixed to get and are fixed on sulfydryl on nano Au particle
The probe P3 of modification.
Above-mentioned preparation method, it is preferred that the ultra-pure water, chlorauric acid solution, sodium citrate solution and ice sodium borohydride
The mass ratio of solution is 9.2: 0.0005~0.0011: 0.0007: 0.0011, and the probe P3 is dense with nanogold particle solution
Degree is than being 2: 0.001~0.002.
Above-mentioned preparation method, it is preferred that in the S1 steps, the carbon doping class graphene phase carbon nitride is using following
Method is prepared：React 24h~25h at being 200 DEG C~220 DEG C in temperature by melamine and absolute ethyl alcohol, then more than
Lifting speed is that 2 DEG C/min~3 DEG C/min is warming up to 520 DEG C~530 DEG C, and 4h~4.5h is calcined at 520 DEG C~530 DEG C, is obtained
To carbon doping class graphene phase carbon nitride.
Above-mentioned preparation method, it is preferred that in the S2 steps, the electro-deposition concretely comprises the following steps：By mass fraction
For 1% HAuCl4The aqueous solution is mixed to get mixed solution with perchloric acid, the glass that carbon doping class graphene phase carbon nitride is modified
Carbon electrode is put into progress current versus time curve scanning in the mixed solution, and the initial potential of electrochemical deposition method is 0V, sampling
At intervals of 0.1s, time 30s~100s, the glass-carbon electrode of nanogold/carbon doping class graphene phase carbon nitride modification is obtained.
Above-mentioned preparation method, it is preferred that the S3 steps are specially：Nanogold/carbon doping class graphene is mutually nitrogenized
The glass-carbon electrode insertion concentration of carbon modification is in 1.0 μM of probe P1, and probe P1 is by chemistry with Electrostatic Absorption in nano-scale gold particle
On son；It is inserted into the mercaptoethanol solution that concentration is 2.0mM, mercaptoethanol is closed nanogold not to be adsorbed, obtain
Probe P1/ nanogold/carbon doping class graphene phase carbon nitride modified glassy carbon electrode.
The technical concept total as one, present invention also offers a kind of above-mentioned electrochemical sensor or above-mentioned preparation method
Application of the electrochemical sensor being prepared in mercury ion is detected.
The application process is：
(1) probe P1/ nanogold/carbon doping class graphene phase carbon nitride modified glassy carbon electrode is soaked in mercury and probe P2
In mixed solution, probe P1, mercury and probe P2 is carried out thymidine-mercury-thymidine complementary pairing, obtain probe P2/ spies
Pin P1/ nanogold/carbon doping class graphene phase carbon nitride modified glassy carbon electrode；
(2) by the probe P2/ probes P1/ nanogold/carbon doping class graphene phase carbon nitride modified glassy carbon electrode and admittedly
The probe P3 reactions being scheduled on nanogold particle, make the probe P3 form double-strand with the probe P1 and probe P2 respectively, obtain
To probe P3/ probe P2/ probes P1/ nanogold/carbon doping class graphene phase carbon nitride modified glassy carbon electrode；
(3) the probe P3/ probes P2/ probes P1/ nanogold/carbon doping class graphene phase carbon nitride is modified into glass carbon electricity
Pole, which is soaked in signal designation agent, reacts；Clean and be soaked in again in 10mM Tris-HCl buffer solutions after drying,
(4) access in the electrolytic cell of three-electrode system and detect current value, line is established according to the concentration of lead ion and current value
Property regression equation：
Y=(2.288 ± 0.0.085) χ+(1.072 ± 0.042)
Wherein, current average when Y is to mercury ion detecting, unit is μ A；χ is the dense of mercury ion in solution to be measured
Degree, the concentration unit of mercury ion is nM；R2For 0.993, detection range is 0.01~1nM, and minimal detectable concentration is 3.7 × 10- 11M。
Above-mentioned application, it is preferred that in (1) step, the probe P1, mercury and probe P2 reaction time be 10~
50min。
Above-mentioned application, it is preferred that in (2) step, the probe P2/ probes P1/ nanogold/carbon doping class graphite
Alkene phase carbon nitride modified glassy carbon electrode and the temperature for the probe P3 reactions being fixed on nanogold particle are 37 DEG C, and the reaction time is
30~90min.
Above-mentioned application, it is preferred that in (3) step, the probe P3/ probes P2/ probes P1/ nanogold/carbon is mixed
It is 10~30min that miscellany graphene phase carbon nitride modified glassy carbon electrode, which is soaked in the reaction time in signal designation agent,.
Above-mentioned application, it is preferred that molten as electrolyte by 5.5~9.0 Tris-HCl of pH in (4) step
Liquid.
Compared with prior art, the advantage of the invention is that：
(1) the invention provides a kind of electrochemical sensor for being used to detect mercury, mutually nitrogenized using carbon doping class graphene
Carbon and nano Au particle modified glassy carbon electrode, repaiied with nanogold/carbon doping class graphene phase carbon nitride multilayer material combination
Decorations detect end surfaces in glass-carbon electrode, optimize the microstructure of reaction end surfaces.Wherein carbon doping class graphene phase carbon nitride
It is the nano material of first stability, there is good dispersiveness and biocompatibility, can effectively maintain bioactivity, makes letter
Just and low cost and other advantages, it is that a kind of outstanding electric signal transmits medium；Nano Au particle has good affine energy to biomolecule
Power and biocompatibility, the transfer velocity of electronics between biology sensor and solution to be measured can be significantly improved, can quickly obtained
Stable response current.The electrochemical sensor of the present invention utilizes the characteristic that has of material in itself, it is possible to increase detection it is efficient
Property and sensitivity.The present invention has taken into full account carbon doping class graphene phase carbon nitride, nanogold, methylene blue and DNA each
Property, and the composite membrane formed using them has highly sensitive, quick response, high measurement accuracy and compared with common-path interference etc.
Characteristic.
(2) the invention provides a kind of electrochemical sensor for being used to detect mercury, P1 and P2 is closed using probe P3, produced
Hybridization reaction, improve current signal；It is due to that nano Au particle has excellent electric conductivity, energy that P3, which is fixed on nano Au particle,
Enough electric signals by hybridization reaction strengthen.
(3) the invention provides a kind of preparation method for being used to detect the electrochemical sensor of mercury, technique is simple, it is fast to make
Speed, and make fixed DNA probe that there is the holding of more preferable stability and high activity.
(4) application of a kind of electrochemical sensor provided by the invention in mercury ion is detected, it is easy to operate, efficiently, inspection
It is low to survey cost, a kind of effective molecular biology for detection is provided for the monitoring and control process of Mercury in Water Body.
Brief description of the drawings
To make the purpose, technical scheme and advantage of the embodiment of the present invention clearer, below in conjunction with the embodiment of the present invention
In accompanying drawing, clear, complete description is carried out to the technical scheme in the embodiment of the present invention.
Fig. 1 is the structure and preparation flow figure of the electrochemical sensor of embodiment 1.
Fig. 2 is the scanning electricity that the glass-carbon electrode that carbon doping class graphene phase carbon nitride is modified in embodiment 2 detects end surfaces
Mirror figure.
Fig. 3 is that the glass-carbon electrode that carbon doping class graphene phase carbon nitride is modified in embodiment 2 detects the carbon content of end surfaces
Energy spectrum diagram.
Fig. 4 is that the glass-carbon electrode of nanogold in embodiment 2/carbon doping class graphene phase carbon nitride modification detects end surfaces
Scanning electron microscope (SEM) photograph.
Fig. 5 is the curent change curve map obtained in embodiment 3 with the mercury of square wave voltammetry detection various concentrations.
Fig. 6 is the linear regression graph of mercury content and curent change in embodiment 3.
Fig. 7 is the selective comparison diagram of electrochemical sensor in embodiment 3.
Embodiment
Below in conjunction with Figure of description and specific preferred embodiment, the invention will be further described, but not therefore and
Limit the scope of the invention.
Material and instrument employed in following examples are commercially available.
Embodiment 1
A kind of electrochemical sensor for being used to detect mercury of the invention, referring to Fig. 1：It is used as including one in three-electrode system
The glass-carbon electrode of working electrode, the test side surface modification of glass-carbon electrode have carbon doping class graphene phase carbon nitride, carbon doping class
Deposition has nano Au particle on graphene phase carbon nitride.Due to being available for adsorption site on nano Au particle, in glass-carbon electrode
The upper probe P1 for adding sulfydryl modification, sulfydryl are adsorbed on the site of nano Au particle, then add mercaptoethanol, close nanometer
The site do not adsorbed in gold particle by sulfydryl.Probe P2 is subsequently added into, probe P2 parts deoxyribonucleotide can be with the mercapto
The probe P1 part deoxyribonucleotides of base modification form duplex structure by thymidine-mercury-thymidine complementary pairing,
In probe P1 and probe P2 the unpaired deoxyribonucleotide of another part all respectively be fixed on it is another on nanogold particle
One is fixed on the probe P3 of sulfydryl modification on nano Au particle, and by base pair complementarity, (fixed P3 nano Au particle is to utilize
The nanogold particle that chemical reduction method makes).
Wherein, probe P1 is preferably with the deoxyribonucleotide sequence shown in SEQ ID NO.1, is specially：
5’-SH-CTGTTTCGTTCCCGAAAGAGGAAG-3’；
Probe P2 is preferably with the nucleotide sequence shown in SEQ ID NO.2, is specially：
5’-AAAGAGGAAGGCCCTTCGTTTCTG-3’；
Probe P3 is preferably with the nucleotide sequence shown in SEQ ID NO.3, is specially：
5’-SH-CTTCCTCTTT-3’
In the presence of having mercury ion in detected water body, because mercury ion and thymidine adhesion in probe are stronger,
Make probe P1 and probe P2 that hybridization reaction occur, form DNA double helical structure, i.e.,：Upper 5 ' the ends of its middle probe P1
" TTCGTTTCTG " at " CTGTTTCGTT " 3 ' end upper with probe P2 carries out complementary pairing.And probe P1 in probe P2 with also existing
A part of unpaired deoxyribonucleotide, now probe P3 and respectively " AAAGAGGAAG " and P2 at 3 ' end upper with probe P1
" AAAGAGGAAG " at upper 5 ' end carries out complementary pairing, forms DNA double helical structure.
Methylene blue is embedded in the gap of DNA double helical structure.DNA generates curent change during forming double-strand,
Nanogold particle expands the change of electric current, methylene blue electric signal indicator the most simultaneously.Due to curent change size with
DNA double chain concentration is relevant, and DNA double chain concentration is relevant with the concentration of mercury ion, the principle system of the invention according to curent change
Standby electrochemical sensor, can effective detection Mercury in Water Body ion concentration.
Embodiment 2
A kind of preparation method of the electrochemical sensor of embodiment 1, specifically includes following steps：
(1) carbon doping class graphene phase carbon nitride is prepared：
1g melamines are placed in the reactor that volume is 100mL, inject absolute ethyl alcohol 80mL, be 200 DEG C in temperature
Lower reaction 24h obtains water white transparency product.After water white transparency product is dried in room temperature, wrapped up with tinfoil, be transferred to Muffle
In stove, 520 DEG C are warming up to by 2 DEG C/min of the rate of climb, and 4h is calcined in 520 DEG C, carbon doping class graphene is obtained and mutually nitrogenizes
Carbon.
(2) pretreatment of glassy carbon electrode：
Glass-carbon electrode (GCE) surface is polished, then rinses glassy carbon electrode surface with water, then uses nitric acid, acetone successively, surpass
Pure water is cleaned by ultrasonic, and it is 7.4 finally to use pH again, and concentration is 10mM Tris-HCl buffer solutions (in Tris-HCl buffer solutions
KCl's containing 1.0M) flushing, naturally dry, obtain pretreated glass-carbon electrode.
(3) preparation of the glass-carbon electrode of carbon doping class graphene phase carbon nitride modification：
3.1st, carbon doping class graphene phase carbon nitride prepared by step (1) is dissolved in ultrasonic 10h, Ran Houli in ultra-pure water
The heart, filtering, 24h is dried in vacuo in being 60 DEG C in temperature, the laminar carbon doping class graphene phase carbon nitride after being peeled off.Carbon
It is block structure to adulterate after class graphene phase carbon nitride is calcined, and electric conductivity is bad, and the laminated structure electric conductivity after stripping carries significantly
It is high.
3.2nd, laminar carbon doping class graphene phase carbon nitride is put into DMF and is prepared into concentration and is
0.05mg/mL suspension, then hanging drop is added to the detection end surfaces of glass-carbon electrode, air-dry at normal temperatures, obtain carbon
Adulterate the glass-carbon electrode of class graphene phase carbon nitride modification.
(4) electro-deposition nano Au particle：
The HAuCl that mass fraction toward 5mL is 1%4The perchloric acid that 200 μ L are added in the aqueous solution is made into mixed solution, will
The glass-carbon electrode of carbon doping class graphene phase carbon nitride modification, which is put into foregoing mixed solution, carries out current versus time curve (I-
T) method scans, and the initial potential of electrochemical deposition method is 0V, sampling interval 0.1s, 30~100s of time, obtains nanogold/carbon
The glass-carbon electrode of class graphene phase carbon nitride modification is adulterated, is dried standby.
(5) probe P1 is modified：
(the probe P1 for being 1.0 μM by the glass-carbon electrode insertion concentration of nanogold/carbon doping class graphene phase carbon nitride modification
It is the probe P1 of sulfydryl modification) in probe P1 (probe P1 is the deoxyribonucleotide sequence shown in SEQ ID NO.1),
Probe P1 is by chemistry and Electrostatic Absorption on nano Au particle；It is inserted into the mercaptoethanol solution that concentration is 2.0mM, makes
Mercaptoethanol closes nanogold not to be adsorbed, obtains probe P1/ nanogold/carbon doping class graphene phase carbon nitride modification glass carbon
Electrode.
(6) chlorauric acid solution and sodium citrate solution are separately added into ultra-pure water, adds ice sodium borohydride while stirring
Dark place reaction is placed in after solution.Ultra-pure water, chlorauric acid solution, the mass ratio of sodium citrate solution and ice sodium borohydride solution are
9.2: 0.0010: 0.0007: 0.0011, obtain nanogold particle solution.Probe P3 is mixed with nanogold particle solution, its is dense
Degree is than being 2: 0.025；Complete the preparation of electrochemical sensor.
Respectively by the glass-carbon electrode and step 4 of the carbon doping class graphene phase carbon nitride being prepared in step 3 modification
The glass-carbon electrode that the nanogold being prepared/have carbon doping class graphene phase carbon nitride is modified carries out Electronic Speculum and energy-spectrum scanning, sweeps
Result is retouched referring to Fig. 2,3,4.
It can be seen that from Fig. 2,3：Glass-carbon electrode test side surface modification has a carbon doping class graphene phase carbon nitride, carbon into
Work(is doped in graphene phase carbon nitride (Fig. 2,3)；
Golden nanometer particle is electrodeposited in the detection end surfaces (figure of the glass-carbon electrode of carbon doping class graphene phase carbon nitride modification
4)。
Embodiment 3
A kind of application of electrochemical sensor of embodiment 1 in mercury ion is detected, specific detection method are：
(1) by probe P2, (ion concentration of mercury is respectively 10 with the mercury ion of gradient concentration respectively-7M、5.0×10-8M、1.0
×10-8M、5.0×10-9M、1.0×10-9M、5.0×10-10M、1.0×10-10M、5.0×10-11M、2.0×10-11M、1.0×
10-11M) it is well mixed, probe P2 concentration is 1 μM.
(2) probe P1/ nanogold/carbon doping class graphene phase carbon nitride modified glassy carbon electrode is soaked in mercury and mixed with probe P2
Close in solution, spy P1, mercury and probe P2 is reacted at normal temperatures 30min.
(3) glass-carbon electrode obtained in step (2) is reacted with being fixed on the P3 on nanogold particle surface at 37 DEG C
60min (reaction time is that 30~90min can be implemented), its middle probe P3 concentration is 2 μM.
(4) glass-carbon electrode of the electrochemical sensor assembled obtained in step (3) is soaked in methylene blue solution
(concentration of this methylene blue is 2 × 10-5M) fully reaction, time are 15min (reaction time is that 10~30min can be implemented),
Clean and be soaked in 15min in 10mM Tris-HCl buffer solutions again after drying.(reaction time is that 10~30min can be implemented),
Then it will access again in the electrolytic cell of three-electrode system, electrolyte solution, detection electric current are used as using the Tris-HCl that pH is 7.4
Value.
Fig. 5 is that ion concentration of mercury is respectively 10-7M(a)、5.0×10-8M(b)、1.0×10-8M(c)、5.0×10-9M(d)、
1.0×10-9M(e)、5.0×10-10M(f)、1.0×10-10M(g)、5.0×10-11M(h)、2.0×10-11M(i)、1.0×10-11The square wave volt-ampere curve (SWV curves) of M (j) solution to be measured.Fig. 6 is the linear regression side of ion concentration of mercury and curent change
Cheng Tu.It was found from from Fig. 5 and Fig. 6, the concentration of mercury ion and the equation of linear regression of current value are：
Y=(2.288 ± 0.0.085) χ+(1.072 ± 0.042)
Wherein, current average when Y is to mercury ion detecting, unit is μ A；χ is the dense of mercury ion in solution to be measured
Degree, the concentration unit of mercury ion is nM；R2For 0.993, detection range is 0.01~1nM, and minimal detectable concentration is 3.7 × 10- 11M。
During aforementioned applications, electrolyte solution is preferably pH 5.5~9.0 Tris-HCl buffer solutions in electrolytic cell.
It can be seen from Fig. 5 and Fig. 6：The minimum detectable concentration of electrochemical sensor of embodiment 1 is 1.0 × 10-14M mercury is molten
Liquid, sensitivity is high, and detection accuracy is high.
Experiment 1：
, now will originally in order to further verify the Detection results of the electrochemical sensor of the present embodiment and its detection method
Water, high mountain foot of a hill or mountain mountain spring water, the solution to be measured of Xiang River water are entered with the electrochemical sensor of embodiment 1 (assay method is with reference to embodiment 3)
The row rate of recovery is tested.
The pretreatment of solution to be measured：Environmental water sample sample is taken, centrifuges filtrate under conditions of 10000r/min after filtering
5min, supernatant liquid filtering is taken to obtain solution to be measured.
Detecting step：Solution to be measured is detected using the electrochemical sensor of embodiment 1.
1 μM of probe P2 and mercury mixed solution are added drop-wise to the reaction end surfaces of the glass-carbon electrode of electrochemical sensor, room temperature
Lower reaction 30 minutes, then the probe P3 (needing to be fixed with nanogold particle) that concentration is 2 μM is dropped in the reaction of glass-carbon electrode
End surfaces, 60min is reacted at 37 DEG C；Then the glass-carbon electrode of the electrochemical sensor assembled is soaked in methylene blue solution
Fully reaction 15min, cleans and is soaked in again in 10mM Tris-HCl buffer solutions after drying, then access the electrode systems of 20mL tri- again
In the electrolytic cell of system, electrolyte solution, detection current value are used as using the Tris-HCl that pH is 7.4.Believed according to caused electric current
Number power correspond to the concentration and content of mercury ion in environmental water sample to be detected.Testing result is as shown in table 1.
Table 1：The rate of recovery the result of three groups of environmental samplesa
| Testing sample molL-1 | Add mercury concentration nmolL-1 | Testing result (10) | The rate of recovery (%) |
| Running water | 10 | 98.58±0.24 | 98.7 |
| High mountain foot of a hill or mountain mountain spring water | 50 | 50.04±0.12 | 100.3 |
| Xiang River water | 100 | 104.01±0.31 | 106.3 |
Note：A represents the mean concentration of measurement.
As can be known from the results of Table 1, mercury ion is detected using the electrochemical sensor of embodiment 1, the rate of recovery is high, has higher
Accuracy.
Experiment 2：
All it is now 20nM K by concentration to verify the electrochemical sensor of the present embodiment selectivity+、Ca2+、Mg2+、Al3 +、Zn2+、Fe3+、Cu2+、Pb2+、Cd2+、Cr2+、Ni+With Hg2+The electrochemical sensor of metal ion solution embodiment 1 to be measured enters
Row measure.
Detecting step：Referring to embodiment 3.
Testing result is as shown in fig. 7, as can be known from Fig. 7：The electrochemical sensor of the present invention is to mercury from higher choosing
Selecting property, antijamming capability rifle are strong.
Said determination result shows, high sensitivity of the present invention, and selectivity is good, and stability is good, can efficiently, low cost online
Lead content is measured, technical support is provided for the monitoring and control process of Mercury in Water Body.
The above described is only a preferred embodiment of the present invention, any formal limitation not is made to the present invention.Though
So the present invention is disclosed as above with preferred embodiment, but is not limited to the present invention.It is any to be familiar with those skilled in the art
Member, in the case where not departing from the Spirit Essence of the present invention and technical scheme, all using in the methods and techniques of the disclosure above
Appearance makes many possible changes and modifications to technical solution of the present invention, or is revised as the equivalent embodiment of equivalent variations.Therefore,
Every content without departing from technical solution of the present invention, the technical spirit according to the present invention is to made for any of the above embodiments any simple
Modification, equivalent substitution, equivalence changes and modification, still fall within technical solution of the present invention protection in the range of.
Claims (10)
1. a kind of electrochemical sensor for being used to detect mercury, it is characterised in that be used as work electricity in three-electrode system including one
The glass-carbon electrode of pole, the test side surface modification of the glass-carbon electrode have carbon doping class graphene phase carbon nitride, the carbon doping
Deposition has nano Au particle on class graphene phase carbon nitride, and the probe P1 of sulfydryl modification is connected on the nano Au particle, institute
Stating electrochemical sensor also includes probe P2 and is fixed on the probe P3 of sulfydryl modification on nano Au particle, the probe P2 parts
Deoxyribonucleotide can pass through thymidine-mercury-thymus gland with the probe P1 part deoxyribonucleotides of the sulfydryl modification
Pyrimidine complementary pairing forms duplex structure, and unpaired deoxyribonucleotide can be with being fixed in the probe P1 and probe P2
The probe P3 of sulfydryl modification forms duplex structure by base pair complementarity on nano Au particle.
2. electrochemical sensor according to claim 1, it is characterised in that the probe P1 is with SEQ ID NO.1
Shown nucleotide sequence；The probe P2 is the nucleotide sequence shown in SEQ ID NO.2；The probe P3 be with
Nucleotide sequence shown in SEQ ID NO.3.
3. electrochemical sensor according to claim 1, it is characterised in that also refer to including signal designation agent, the signal
Show the duplex structure that agent is embedded in the duplex structure that the probe P1 and probe P2 is formed, probe P1 and probe P3 is formed, and
In the duplex structure that probe P2 and probe P3 is formed.
4. electrochemical sensor according to claim 1, it is characterised in that the signal designation agent is methylene blue, institute
It is 0.1~0.5mM to state methylene blue concentration.
5. the preparation method of electrochemical sensor any one of a kind of Claims 1-4, it is characterised in that including following
Step：
S1, carbon doping class graphene phase carbon nitride is configured to suspension, the detection end surfaces for being added drop-wise to glass-carbon electrode obtain carbon
Adulterate the glass-carbon electrode of class graphene phase carbon nitride modification；
S2, nano Au particle is electrodeposited in carbon doping class graphene phase carbon nitride modification glass-carbon electrode test side table
Face obtains the glass-carbon electrode of nanogold/carbon doping class graphene phase carbon nitride modification；
S3, the glass-carbon electrode for modifying nanogold/carbon doping class graphene phase carbon nitride are inserted in the probe P1 of sulfydryl modification, institute
The probe P1 of sulfydryl modification is stated by Electrostatic Absorption on nano Au particle；It is inserted into mercaptoethanol solution, makes sulfydryl second
Alcohol closes nanogold not to be adsorbed, obtains probe P1/ nanogold/carbon doping class graphene phase carbon nitride modified glassy carbon electrode.
6. preparation method according to claim 5, it is characterised in that also include with manufacturing probe P2 solution and be fixed on nanometer
The probe P3 solution of sulfydryl modification in gold particle；The probe P3 solution of sulfydryl modification on nano Au particle is fixed on described in preparation
Concretely comprise the following steps：Ultra-pure water, chlorauric acid solution and sodium citrate solution are mixed, after adding ice sodium borohydride solution while stirring
Dark place is placed in react to obtain nanogold particle solution；Probe P3 and the nanogold particle solution are mixed to get and are fixed on nanometer
The probe P3 of sulfydryl modification in gold particle.
7. preparation method according to claim 6, it is characterised in that the ultra-pure water, chlorauric acid solution, sodium citrate are molten
The mass ratio of liquid and ice sodium borohydride solution is 9.2: 0.0005~0.0011: 0.0007: 0.0011, the probe P3 and nanometer
The concentration ratio of gold grain solution is 2: 0.001~0.002.
8. the preparation method according to any one of claim 5 to 7, it is characterised in that
In the S1 steps, the carbon doping class graphene phase carbon nitride is prepared using following methods：By melamine and
Absolute ethyl alcohol reacts 24h at being 200 DEG C in temperature, is then warming up to 520 DEG C by 2 DEG C/min of the rate of climb, and forge in 520 DEG C
4h is burnt, obtains carbon doping class graphene phase carbon nitride；
And/or in the S1 steps, carbon doping class graphene phase carbon nitride is configured to concretely comprising the following steps for suspension：By carbon
Adulterate class graphene phase carbon nitride and peel off flakiness shape carbon doping class graphene phase carbon nitride, by laminar carbon doping class graphene
Phase carbon nitride, which is scattered in DMF, is made suspension；
And/or in the S2 steps, the electro-deposition concretely comprises the following steps：By the HAuCl that mass fraction is 1%4The aqueous solution with
Perchloric acid is mixed to get mixed solution, and the glass-carbon electrode that carbon doping class graphene phase carbon nitride is modified is put into the mixed solution
Middle progress current versus time curve scanning, the initial potential of electrochemical deposition method are 0V, sampling interval 0.1s, the time 30~
100s, obtain the glass-carbon electrode of nanogold/carbon doping class graphene phase carbon nitride modification；
And/or the S3 steps are specially：The glass-carbon electrode of nanogold/carbon doping class graphene phase carbon nitride modification is inserted
Concentration is in 1.0 μM of probe P1, and probe P1 is by chemistry and Electrostatic Absorption on nano Au particle；Being inserted into concentration is
In 2.0mM mercaptoethanol solution, mercaptoethanol is closed nanogold not to be adsorbed, obtain probe P1/ nanogold/carbon doping
Class graphene phase carbon nitride modified glassy carbon electrode.
9. preparation any one of electrochemical sensor any one of a kind of Claims 1-4 or claim 5 to 8
Application of the electrochemical sensor that method is prepared in mercury ion is detected, it is characterised in that the application process is：
(1) probe P1/ nanogold/carbon doping class graphene phase carbon nitride modified glassy carbon electrode is soaked in mercury and mixed with probe P2
In solution, probe P 1, mercury and probe P2 is carried out thymidine-mercury-thymidine complementary pairing, obtain probe P2/ probes
P1/ nanogold/carbon doping class graphene phase carbon nitride modified glassy carbon electrode；
(2) by the probe P2/ probes P1/ nanogold/carbon doping class graphene phase carbon nitride modified glassy carbon electrode with being fixed on
Probe P3 reactions on nanogold particle, make the probe P3 form double-strand with the probe P1 and probe P2 respectively, are visited
Pin P3/ probe P2/ probes P1/ nanogold/carbon doping class graphene phase carbon nitride modified glassy carbon electrode；
(3) the probe P3/ probes P2/ probes P1/ nanogold/carbon doping class graphene phase carbon nitride modified glassy carbon electrode is soaked
Steep and reacted in signal designation agent；Clean the Tris-HCl buffer solutions for being soaked in 10mM after drying again；
(4) access in the electrolytic cell of three-electrode system and detect current value, linear return is established according to the concentration of lead ion and current value
Return equation：
Y=(2.288 ± 0.0.085) χ+(1.072 ± 0.042)
Wherein, current average when Y is to mercury ion detecting, unit is μ A；χ be solution to be measured in mercury ion concentration, mercury
The concentration unit of ion is nM；R2For 0.993, detection range is 0.01~1nM, and minimal detectable concentration is 3.7 × 10-11M。
10. application according to claim 9, it is characterised in that in (1) step, probe P 1, mercury and the probe
The time of P2 reactions is 10~50min；
And/or in (2) step, the probe P2/ probes P1/ nanogold/carbon doping class graphene phase carbon nitride modification glass
Carbon electrode and the temperature for the probe P3 reactions being fixed on nanogold particle are 37 DEG C, and the reaction time is 30~90min；
And/or in (3) step, the probe P3/ probes P2/ probes P1/ nanogold/carbon doping class graphene mutually nitrogenizes
It is 10~30min that carbon modified glassy carbon electrode, which is soaked in the time reacted in signal designation agent,；
And/or in (4) step, electrolyte solution is used as by 5.5~9.0 Tris-HCl of pH.
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