CN109187689A - A kind of self assembly electrochemical sensor and its application in water phase in the detection of trace amount mercury ion - Google Patents
A kind of self assembly electrochemical sensor and its application in water phase in the detection of trace amount mercury ion Download PDFInfo
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Abstract
The invention discloses a kind of self assembly electrochemical sensor and its applications in water phase in the detection of trace amount mercury ion.The self assembly electrochemical sensor is three-electrode system electrochemical sensor, using saturated calomel electrode as reference electrode, platinum electrode is auxiliary electrode, using above-mentioned mercaptoethylmaine/thymidine -1- acetic acid gold electrode as working electrode, preenrichment is carried out to mercury ion using chronoamperometry, current value corresponding to mercury ion under various concentration is recorded with differential pulse method, draw the working curve of the amperometric sensor of detection mercury ion, and can use working curve method, obtain the concentration of mercury ion in sample to be tested.The present invention uses thymidine -1- acetic acid as capture probe, compared to DNA chain is used as capture probe, is easier to complete in laboratory, preparation is simple, cheap.
Description
Technical field:
The present invention relates to electrochemical analysis techniques fields, and in particular to a kind of self assembly electrochemical sensor and its in water phase
Application in the detection of middle trace amount mercury ion.
Background technique:
Mercury is that have highly toxic global environmental contaminants, and especially it is turned into high migration, persistence, methyl
The characteristics of with property, bioconcentration and food chain amplification, the mercury of even denier exists in the environment, to aquatile and
The health of the mankind also has greatly harm.There is (inorganic, organic or mixed form) in a variety of forms in the environment in mercury, water-soluble
The dimercurion (Hg2+) of property is one of most common and most stable of form of mercury pollution.Mercury is tapped into directly or by between food chain
After entering human body, it can be accumulated in brain, liver and other organs, to other portions of nervous system, digestive system, immune system and body
Position generates serious toxic effect.So the mercury content in the water environment of monitoring analysis with sensitivity, to including human health and life
State environment has a very important significance.
Mercury content method for monitoring and analyzing traditional at present mainly has: inductive coupling plasma mass spectrometry (ICP-MS), atom hair
Penetrate spectrum (AES), atomic fluorescence spectrometry (AFS), chromatography of ions (IC) etc., although these methods can obtain it is more accurate
Testing result, but these methods rely on large-scale instrument and equipment, sample need to carry out that complicated pretreatment, consuming time is long, detection
At high cost and special technical staff is needed to operate, these defects limit conventional method and examine in environmental sample Trace Hg
Application in survey.So the measurement of mercury ion content in environment how is effectively performed, be still vast analysis staff urgently
Problem to be solved.
In recent years, using the heavy metal in electrochemical method analysis detection environment, due to its high specificity, high sensitivity,
Detecting the features such as rapid, low in cost is concerned by people, and becomes an important research hot spot of environmental monitoring.Scientific research personnel
By modifying electrochemical sensor using various new materials to improve the stability, repeatability and structure of electrochemical sensor
Reliability.According to research reports, mercury ion can be with the mercury specific oligonucleotide segment mispairing rich in thymidine (T) in solution
Form T-Hg2+- T structure.In recent years, it is embedded in form T-Hg using double-stranded DNA2+The electrochemical sensor technology of-T structure develops
Rapidly, this kind of sensor shows higher sensitivity and selectivity in the detection of mercury ion.But current business DNA chain at
This is higher, and the activity of DNA is easy to be influenced by environment, limits its application in terms of electrochemical sensor.Researcher
Constantly look for new material instead of business DNA chain to reduce cost, adjust electrochemical sensor structure with improve its stability,
Repeatability, reliability realize simple, sensitive, quick, the inexpensive detection for mercury ion in water.
Summary of the invention:
The inspection of trace amount mercury ion the purpose of the present invention is to provide a kind of self assembly electrochemical sensor and its in water phase
Application in survey, the present invention use thymidine -1- acetic acid as capture probe, compared to use DNA chain as capture probe,
It is easier to complete in laboratory, preparation is simple, cheap.
The present invention is achieved by the following technical programs:
The present invention provides a kind of mercaptoethylmaine/thymidine -1- acetic acid gold electrodes, are prepared by following steps:
(1) the Au electrode after polishing and cleaning is immersed in the mixed solution of the second alcohol and water of mercaptoethylmaine and is impregnated
8~10h, soaking temperature are 4 DEG C~20 DEG C, the concentration of mercaptoethylmaine in the mixed solution of the second alcohol and water of the mercaptoethylmaine
For 1.5~2.5mg/mL；
(2) it will be soaked in thymidine -1- acetic acid mixed solution in step (1) by the Au electrode that mercaptoethylmaine impregnated
Steep 8~10h, soaking temperature be 4 DEG C~20 DEG C to get arrive working electrode, the thymidine -1- acetic acid mixed solution mesothorax
The concentration of gland pyrimidine -1- acetic acid is 0.5~1.5mg/mL.Mercaptoethylmaine proposed by the present invention/thymidine -1- acetic acid gold electrode
Preparation method, simple process, environmentally protective, the mercaptoethylmaine being prepared/thymidine -1- acetic acid gold electrode realizes pair
The quantitative detection of trace amount mercury ion in water phase.
The present invention, as substrate, ensure that the abundant use of electrode surface, whole cross section is equal using commercialization gold electrode
It can be self-assembly of Au-S key with mercaptoethylmaine, provide wide platform for the link of capture probe；Meanwhile gold electrode is compared
In the glass-carbon electrode of the modifications such as graphene, being bonded between electrode and capture probe is stronger, thus ensure that Trace Hg from
The stability of son detection, repeatability.Using thymidine as capture interface, formed using thymidine and mercury ion special
The T-Hg of property2+- T structure avoids other impurities heavy metal ion in water phase, such as Cd2+, Pb2+, Cu2+, Ni2+, Zn2+, Co2+'s
Interference, improves the specificity to mercury ion detecting.
It is preferred that Au electrode described in step (1) is comprised the concrete steps that by polishing and cleaning treatment: by diameter be 3~
The Au electrode of 5mm successively uses the Al of 300nm and 50nm2O3Powder polishing, is successively cleaned up with dehydrated alcohol and ultrapure water.
It is preferred that thymidine -1- acetic acid mixed solution described in step (2) is prepared by following steps: in concentration
It is sub- for addition N- hydroxysuccinimidyl acyl in 2- (N- morpholine) the ethanesulfonic acid buffer solution (MES buffer solution) of 50~150mmol/L
Amine (NHS) and 1- (3- dimethylaminopropyl) -3- ethyl-carbodiimide hydrochloride (EDC) are configured to n-hydroxysuccinimide
With 2- (N- morpholine) ethanesulfonic acid buffer solution of 1- (3- dimethylaminopropyl) -3- ethyl-carbodiimide hydrochloride, N- hydroxyl
The concentration of base succinimide is 50~150mmol/L, 1- (3- dimethylaminopropyl) -3- ethyl-carbodiimide hydrochloride
Concentration is 50~150mmol/L, to n-hydroxysuccinimide and 1- (3- dimethylaminopropyl) -3- ethyl carbodiimide salt
Thymidine -1- acetic acid is added in 2- (N- morpholine) ethanesulfonic acid buffer solution of hydrochlorate, the concentration of thymidine -1- acetic acid is
0.5~1.5mg/mL.The concentration of MES buffer solution is 50~150mmol/L, pH=6.
It is preferred that the volume ratio of ethyl alcohol described in step (1) and water is 3:1.
Another object of the present invention is to provide a kind of self assembly electrochemical sensor, which is three electricity
Polar body system electrochemical sensor, using saturated calomel electrode as reference electrode, platinum electrode is auxiliary electrode, with above-mentioned sulfydryl second
Amine/thymidine -1- acetic acid gold electrode is working electrode.
The present invention also provides application of the above-mentioned self assembly electrochemical sensor in water phase in the detection of trace amount mercury ion.
It is preferred that the detection of self assembly electrochemical sensor trace amount mercury ion in water phase, using three-electrode system, with saturation
Calomel electrode is reference electrode, and platinum electrode is auxiliary electrode, using mercaptoethylmaine/thymidine -1- acetic acid gold electrode as work
Electrode, electrolyte are the mercury ion solution that pH is 5~8, are included the following steps, carry out pre- richness to mercury ion using chronoamperometry
Collection, setting voltage be -1.0~0.1V, 0.1~0.5s of sampling interval, 200~300s of runing time, using differential pulse method into
Row detection, the parameter of differential pulse method are as follows: take-off potential is -1.0V, and termination current potential is 0.6V, 10~50s of quiescent time, pulse
Amplitude be 0.02~0.06V, pulse increment be 0.01~0.02V, pulse width be 0.02~0.1s, the pulse period be 0.05~
0.5s draws the working curve of the electrochemical sensor of detection mercury ion, using working curve method obtain in sample to be tested mercury from
The concentration of son.
The working curve is the linear relationship curve of current value and ion concentration of mercury, current value and ion concentration of mercury
Linear relationship such as following formula indicates: y=0.1271x+8.4551, in which: current value when y is to mercury ion detecting, unit are μ A,
X is the concentration of mercury ion in solution to be measured, unit nmol/L, coefficient R2It is 0.99857.
The action principle of self assembly electrochemical sensor proposed by the present invention are as follows: access prepared electrochemical sensor
Three-electrode system simultaneously carries out electro-chemical test as working electrode, is platinum electrode to electrode, and reference electrode is saturation calomel electricity
Pole, test environment are normal temperature and pressure.Electrolyte selects 10mL, 10nmol/L, 50nmol/L, 100nmol/L, 200nmol/ respectively
L, the mercury ion mixed solution that the pH of 300nmol/L is 5~8, the pH of electrolyte are adjusted with the buffer of 0.5mol/L PBS
Section.Preenrichment is carried out to mercury ion using chronoamperometry, differential pulse genetic method is reused and is detected, record various concentration
Current value corresponding to mercury ion is drawn the working curve of the amperometric sensor of detection mercury ion, is obtained using working curve method
The concentration of mercury ion into sample to be tested.
The beneficial effects of the present invention are:
(1) present invention uses thymidine -1- acetic acid as capture probe, compared to use DNA chain as capture probe,
It is easier to complete in laboratory, preparation is simple, cheap.
(2) present invention forms the T- of specificity using thymidine and mercury ion using thymidine as capture interface
Hg2+- T structure avoids other impurities heavy metal ion in water phase, such as Cd2+, Pb2+, Cu2+, Ni2+, Zn2+, Co2+Interference, mention
The high specificity to mercury ion detecting.
(3) present invention ensure that the abundant use of electrode surface, whole cross section using commercialization gold electrode as substrate
It can be self-assembly of Au-S key with mercaptoethylmaine, provide wide platform for the link of capture probe；Meanwhile gold electrode phase
Than in the glass-carbon electrode of the modifications such as graphene, being bonded between gold electrode and capture probe is stronger, therefore ensure that trace
The stability of mercury ion detecting, repeatability.
Detailed description of the invention:
Fig. 1 is the structure and preparation flow figure of electrochemical sensor of the invention；
Fig. 2 is to be become in the embodiment of the present invention with the voltage-to-current that the mercury ion of differential pulse method detection various concentration obtains
Change curve graph；
Fig. 3 is the linear regression graph of ion concentration of mercury and curent change when carrying out Electrochemical Detection in the embodiment of the present invention.
Specific embodiment:
The following examples are further illustrations of the invention, rather than limiting the invention.
Except special instruction, equipment mentioned in the present invention and material are commercially available.
Embodiment 1:
As shown in Figure 1, mercaptoethylmaine/thymidine -1- acetic acid gold electrode, is prepared by following steps:
(1) gold electrode that diameter is 3mm is successively used into 300nm and 50nm Al2O3It is clear with ethyl alcohol, ultrapure water after powder polishing
Wash clean；
(2) polished gold electrode is immersed in the mixed solution of the second alcohol and water for the mercaptoethylmaine that concentration is 1.5mg/mL
In, wherein the volume ratio of ethyl alcohol and water is 3:1, at 4 DEG C, impregnates 8h；
(3) in the 2- of 50mmol/L (N- morpholine) ethanesulfonic acid buffer solution (MES buffer solution, pH=6), 50mmol/L
N-hydroxysuccinimide (NHS) and 50mmol/L 1- (3- dimethylaminopropyl) -3- ethyl-carbodiimide hydrochloride
(EDC) in mixed solution, appropriate thymidine -1- acetic acid is added, is made into -1 acetic acid solution of thymidine of 0.5mg/mL, and
Electrode after mercaptoethylmaine is impregnated is put into thymidine -1- acetic acid mixed solution, impregnates 8h at 4 DEG C.
Embodiment 2:
As shown in Figure 1, mercaptoethylmaine/thymidine -1- acetic acid gold electrode, is prepared by following steps:
(1) gold electrode that diameter is 4mm is successively used into 300nm and 50nm Al2O3After powder polishing, then with ethyl alcohol, ultrapure water
It cleans up；
(2) polished gold electrode is immersed in the ethyl alcohol of mercaptoethylmaine and the mixed solution of water of 2.0mg/mL,
The volume ratio of middle ethyl alcohol and water is 3:1, at 10 DEG C, impregnates 9h；
(3) in the 2- of 100mmol/L (N- morpholine) ethanesulfonic acid buffer solution (MES buffer solution, pH=6),
The n-hydroxysuccinimide (NHS) of 100mmol/L and 1- (3- the dimethylaminopropyl) -3- ethyl carbon two of 100mmol/L
In the mixed solution of inferior amine salt hydrochlorate (EDC), appropriate thymidine -1- acetic acid is added, is made into the thymidine -1 of 1.0mg/mL
Acetic acid solution, and the electrode after mercaptoethylmaine is impregnated is put into thymidine -1- acetic acid mixed solution, is impregnated at 10 DEG C
9h。
Embodiment 3:
As shown in Figure 1, mercaptoethylmaine/thymidine -1- acetic acid gold electrode, is prepared by following steps:
(1) gold electrode that diameter is 5mm is successively used into 300nm and 50nm Al2O3After powder polishing, then with ethyl alcohol, ultrapure water
It cleans up；
(2) polished gold electrode is immersed in the ethyl alcohol of mercaptoethylmaine and the mixed solution of water of 2.5mg/mL,
The volume ratio of middle ethyl alcohol and water is 3:1, at 20 DEG C, impregnates 10h；
(3) in the 2- of 150mmol/L (N- morpholine) ethanesulfonic acid buffer solution (MES buffer solution, pH=6),
The n-hydroxysuccinimide (NHS) of 150mmol/L and 1- (3- the dimethylaminopropyl) -3- ethyl carbon two of 150mmol/L
In the mixed solution of inferior amine salt hydrochlorate (EDC), appropriate thymidine -1- acetic acid is added, is made into the thymidine -1 of 1.5mg/mL
Acetic acid solution, and the electrode after mercaptoethylmaine is impregnated is put into thymidine -1- acetic acid mixed solution, is impregnated at 20 DEG C
10h。
Embodiment 4:
The mercaptoethylmaine being prepared using embodiment 1/thymidine -1- acetic acid gold electrode carries out water phase as working electrode
The working curve of mercury ion standard solution is drawn in the measurement of trace amount mercury ion, the specific steps are as follows:
(1) it is tested using three-electrode system, saturated calomel electrode is reference electrode, and platinum electrode is to electrode, in fact
Applying mercaptoethylmaine/thymidine -1- acetic acid gold electrode that example 1 is prepared is working electrode, is 5 (use in 10mL pH
0.5mol/LPBS buffer solution adjust pH) various concentration mercury ion solution in tested, the mercury in mercury ion solution from
Sub- concentration is respectively 10nmol/L, 50nmol/L, 100nmol/L, 200nmol/L and 300nmol/L；
(2) be 5 in 10mL pH (adjusting pH with 0.5mol/L PBS buffer solution) ion concentration of mercury for 10nmol/L
Mercury ion solution tested for, first using chronoamperometry to mercury ion carry out preenrichment, setting voltage be-
1.0V, sampling interval 0.1s, runing time 200s；Analyte is detected with differential pulse method again, major parameter are as follows: starting
Current potential is -1.0V, and terminations current potential is 0.6V, quiescent time 10s, impulse amplitude 0.02V, and pulse increment 0.01V, pulse is wide
Degree is 0.02s, pulse period 0.05s；
The test method of the ion concentration of mercury of other mercury ion solution is for example above-mentioned.
(3) current value corresponding to mercury ion, drafting detect the current mode of mercury ion under various concentration in recording step (2)
The working curve (as shown in Figure 2) of sensor.
Data in analysis chart 2 obtain the equation of linear regression of current value and ion concentration of mercury are as follows:
Y=0.1271x+8.4551
Wherein, current value when y is to mercury ion detecting, unit are μ A；X is the concentration of mercury ion in solution to be measured, mercury
The concentration unit of ion is nmol/L；Linear regression curves are as shown in figure 3, the range of linearity of diagramatic curve is 10~300nmol/
L, coefficient R2It is 0.99857.
Embodiment 5:
It is measured with the electrochemical sensor of embodiment 1, unknown water is calculated according to working curve obtained in embodiment 4
The ion concentration of mercury of sample:
(1) a certain amount of environmental water sample is taken, is filtered, filtrate is taken to be centrifuged 6min under the revolving speed of 10000r/min, is protected
Stay supernatant；
(2) appropriate supernatant is taken, PBS buffer solution is added, adjusting pH is 5, adds appropriate NaCl, makes its concentration
0.5mol/L prepares 10ml solution to be measured；
(3) analyte is detected with differential pulse voltammetry, mercury ion is carried out using chronoamperometry first pre-
Enrichment, setting voltage are -1.0V, sampling interval 0.1s, runing time 200s；It is detected again with differential pulse method, it is main to join
Number are as follows: take-off potential is -1.0V, and termination current potential is 0.6V, quiescent time 10s, impulse amplitude 0.02V, and pulse increment is
0.01V, pulse width 0.02s, pulse period 0.05s；
(4) it measures three times, records the peak point current of sample to be tested, be averaged as 13.64 μ A, with the line of working curve
Property regression equation find out ion concentration of mercury be 40.79nM.Identical water sample is detected using atomic fluorescence method, determines that its is dense
Degree is 40.38nM, and method of the invention is compared with other methods, and testing result differs only by 1.02%, electrification proposed by the present invention
It is suitable with other methods to learn sensor test accuracy.
Above to a kind of self assembly electrochemical sensor provided by the invention and its in water phase trace amount mercury ion detection
In application be described in detail, technical solution of the present invention that the above embodiments are only used to help understand and its
Core concept, it is noted that those of ordinary skill in the art, without departing from the principle of the present invention, also
Can be with several improvements and modifications are made to the present invention, these improvement and modification also fall into the protection scope of the claims in the present invention
It is interior.
Claims (8)
1. a kind of mercaptoethylmaine/thymidine -1- acetic acid gold electrode, which is characterized in that be prepared by following steps:
(1) the Au electrode after polishing and cleaning is immersed in the mixed solution of the second alcohol and water of mercaptoethylmaine impregnate 8~
10h, soaking temperature are 4 DEG C~20 DEG C, and the concentration of mercaptoethylmaine is in the mixed solution of the second alcohol and water of the mercaptoethylmaine
1.5~2.5mg/mL；
(2) 8 will be impregnated in thymidine -1- acetic acid mixed solution by the Au electrode that mercaptoethylmaine impregnated in step (1)
~10h, soaking temperature are 4 DEG C~20 DEG C to get arriving working electrode, thymus gland in the thymidine -1- acetic acid mixed solution
The concentration of pyrimidine -1- acetic acid is 0.5~1.5mg/mL.
2. mercaptoethylmaine according to claim 1/thymidine -1- acetic acid gold electrode, which is characterized in that in step (1)
The polishing of Au electrode process and cleaning treatment comprises the concrete steps that: the Au electrode that diameter is 3~5mm is successively used 300nm
With the Al of 50nm2O3Powder polishing, is successively cleaned up with dehydrated alcohol and ultrapure water.
3. mercaptoethylmaine according to claim 1/thymidine -1- acetic acid gold electrode, which is characterized in that in step (2)
Thymidine -1- the acetic acid mixed solution is prepared by following steps: in the 2- (N- that concentration is 50~150mmol/L
Morpholine) n-hydroxysuccinimide and 1- (3- dimethylaminopropyl) -3- ethyl carbon two are added in ethanesulfonic acid buffer solution
Inferior amine salt hydrochlorate is configured to n-hydroxysuccinimide and 1- (3- dimethylaminopropyl) -3- ethyl-carbodiimide hydrochloride
2- (N- morpholine) ethanesulfonic acid buffer solution, the concentration of n-hydroxysuccinimide are 50~150mmol/L, 1- (3- dimethyl
Aminopropyl) -3- ethyl-carbodiimide hydrochloride concentration be 50~150mmol/L, to n-hydroxysuccinimide and 1- (3-
Dimethylaminopropyl) -3- ethyl-carbodiimide hydrochloride 2- (N- morpholine) ethanesulfonic acid buffer solution in be added thymus gland it is phonetic
Pyridine -1- acetic acid, the concentration of thymidine -1- acetic acid are 0.5~1.5mg/mL.
4. mercaptoethylmaine according to claim 1/thymidine -1- acetic acid gold electrode, which is characterized in that in step (1)
The volume ratio of the ethyl alcohol and water is 3:1.
5. a kind of self assembly electrochemical sensor, which is characterized in that the electrochemical sensor is three-electrode system electrochemical sensing
Device, using saturated calomel electrode as reference electrode, platinum electrode is auxiliary electrode, and mercaptoethylmaine/thymus gland that claim 1 obtains is phonetic
Pyridine -1- acetic acid gold electrode is working electrode.
6. application of the self assembly electrochemical sensor in water phase in the detection of trace amount mercury ion described in claim 5.
7. application of the self assembly electrochemical sensor according to claim 6 in water phase in the detection of trace amount mercury ion,
It is characterized in that, using saturated calomel electrode as reference electrode, platinum electrode is auxiliary electrode, with sulfydryl using three-electrode system
Ethamine/thymidine -1- acetic acid gold electrode is working electrode, and electrolyte is the mercury ion solution that pH is 5~8, using timing electricity
Stream method to mercury ion carry out preenrichment, setting voltage be -1.0~0.1V, 0.1~0.5s of sampling interval, runing time 200~
300s is detected using differential pulse method, the parameter of differential pulse method are as follows: take-off potential is -1.0V, terminates current potential and is
0.6V, 10~50s of quiescent time, impulse amplitude are 0.02~0.06V, and pulse increment is 0.01~0.02V, and pulse width is
0.02~0.1s, pulse period are 0.05~0.5s, draw the working curve of the electrochemical sensor of detection mercury ion, utilize work
The concentration of mercury ion in sample to be tested is obtained as curve method.
8. application of the self assembly electrochemical sensor according to claim 7 in water phase in the detection of trace amount mercury ion,
It is characterized in that, the working curve is the linear relationship curve of current value and ion concentration of mercury, current value is dense with mercury ion
The linear relationship of degree such as following formula indicates: y=0.1271x+8.4551, in which: current value when y is to mercury ion detecting, unit
It is the concentration of mercury ion in solution to be measured, unit nmol/L, coefficient R for μ A, x2It is 0.99857.
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|---|---|---|---|---|
| CN110907518A (en) * | 2019-11-29 | 2020-03-24 | 中南大学 | Preparation and detection method of mercury ion electrochemical biosensor |
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