CN104155289B - A kind of solid-state electrochemistry illumination sensor detecting mercury ion and its preparation method and application - Google Patents
A kind of solid-state electrochemistry illumination sensor detecting mercury ion and its preparation method and application Download PDFInfo
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- CN104155289B CN104155289B CN201410289966.1A CN201410289966A CN104155289B CN 104155289 B CN104155289 B CN 104155289B CN 201410289966 A CN201410289966 A CN 201410289966A CN 104155289 B CN104155289 B CN 104155289B
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Abstract
The present invention discloses a kind of solid-state electrochemistry illumination sensor for mercury ion detecting and its preparation method and application, the ABEI of feature to be this sensor be surperficial immobilized electropolymerization, then be assembled with successively glutaraldehyde, DNA1, by T-Hg2+The vitamin H of DNA2, DNA2 end that-T mispairing combines and the glass-carbon electrode of avidin;Its preparation method comprises immobilized has the preparation process of DNA1 glass-carbon electrode and the number of assembling steps of solid-state electrochemistry illumination sensor, advantage be have highly sensitive, good stability, selectivity are strong, circulation ratio is good, be easy to operation, save reagent etc.
Description
Technical field
The present invention relates to heavy metal analysis technical field, especially relate to a kind of solid-state electrochemistry illumination sensor detecting mercury ion and its preparation method and application.
Background technology
Heavy metal contamination is one of important component part of present environmental pollution. Mercury (Hg), is unique liquid metal under normal temperature, is the number one killer in heavy metal contamination. In recent years, the exploitation of mankind's heavy metal mercury, smelting, processing and business manufacturing activities increase day by day, and a large amount of mercury enters in air, water, soil and retains, accumulates and move. Mercury in environment finally enters in the body of animals and humans with the form of food chain, and the vital functions of organism is caused very big injury, produces psychoneural symptom, trembles, the disease such as stomatitis and toxic nephropathy. Mercury ion (Hg2+) be one of main existence form in nature of mercury, in numerous hygienic standard requirement must not detection, environment measuring and food safety monitoring are had important meaning by the lower concentration mercury ion detecting method therefore developing highly sensitive and highly selective.
The method of detection mercury ion mainly contains at present: cold atomic absorption spectrometry, ICP-MS, fluorescent spectrometry, ultraviolet-visible spectrophotometry, atomic fluorescence method, electrochemical process, ion chromatography, capillary electrophoresis, heavy metal rapid detector method, test strip method etc., but these methods maybe need to use expensive instrument, operate loaded down with trivial details, and require that testing staff possesses certain expertise, analysis cost height, it is difficult to universal; Or sensitivity is not high, poor selectivity, sample pretreatment is complicated, it is difficult to accurately detection by quantitative lower concentration Hg2+��
Electrochemiluminescence (ECL) is a kind of analysis technology electrochemistry combined with chemoluminescence, not only there is highly sensitive, the linearity range width of chemiluminometry, observe the advantages such as convenient, instrument is simple, and the current potential controllability with electrochemical methods is strong, selectivity height, stable reagent, high repeatability and other advantages, the very big concern being subject to researcher, and develop into the important branch analyzing detection field gradually. Solid-state electrochemistry illumination is by the immobilized technology carrying out ECL detection after electrode surface again of chemical process or physical method by electrochemiluminescence reagent, compared with the ECL in solution, electrochemiluminescence reagent is fixed to the solid-state ECL that electrode surface builds, also has following advantage: reduce the consumption of expensive reagent, simplify experimental implementation, improve ECL intensity etc. But, at present, any solid-state electrochemistry illumination detection Mercury in Water Body ion content relevant report utilizing N-(4-ammonia butyl) the different luminol,3-aminophthalic acid cyclic hydrazide of-N-ethyl (ABEI) is not also disclosed both at home and abroad.
Summary of the invention
Technical problem to be solved by this invention is to provide that a kind of highly sensitive, good stability, selectivity are strong, favorable reproducibility, the solid-state electrochemistry illumination sensor for mercury ion detecting being easy to operation and its preparation method and detection method.
The present invention solves the problems of the technologies described above the technical scheme adopted: a kind of solid-state electrochemistry illumination sensor for detecting mercury ion, and described sensor is the ABEI of surperficial immobilized electropolymerization, then be assembled with successively glutaraldehyde, DNA1, by T-Hg2+The vitamin H of DNA2, DNA2 end that-T mispairing combines and the glass-carbon electrode of avidin.
The preparation method of the above-mentioned solid-state electrochemistry illumination sensor for detecting mercury ion, concrete steps are as follows:
(1) the immobilized preparation having DNA1 glass-carbon electrode
A. by diameter be the glass-carbon electrode of 3��5mm successively with the aluminium sesquioxide polished finish of 1.0 ��m, 0.3 ��m, 0.05 ��m, then successively with ethanol, water ultrasonic cleaning, after water is rinsed well, nitrogen blows dry for subsequent use;
B. step (a) gained glass-carbon electrode is placed in the H containing N-(4-ammonia butyl) the different luminol,3-aminophthalic acid cyclic hydrazide of-N-ethyl (ABEI)2SO4In solution, carrying out cyclic voltammetry scan, voltage range is-0.2��1.5V, encloses with the speed scan round 20��30 of sweeping of 0.01��0.1V/s, makes ABEI at glassy carbon electrode surface electropolymerization; The described H containing N-(4-ammonia butyl) the different luminol,3-aminophthalic acid cyclic hydrazide of-N-ethyl (ABEI)2SO4In solution, N-(4-ammonia butyl)-N-ethyl different luminol,3-aminophthalic acid cyclic hydrazide (ABEI) concentration is 0.01��0.0001mol/L, H2SO4Concentration is 0.1��1mol/L;
C. after 0.01��0.1MPBS damping fluid of step (b) gained glass-carbon electrode pH=7��8 being rinsed well, drip in glassy carbon electrode surface and add 1��3wt% glutaraldehyde solution 10��20 �� L, leave standstill 20��40 minutes;
D. after 0.01��0.1MPBS damping fluid of step (c) gained glass-carbon electrode pH=7��8 being rinsed well, 0.01��0.1MPBS damping fluid 10��20 �� L of pH=7��8 added containing 10 ��m of ol/LDNA1 is dripped in glassy carbon electrode surface, leave standstill 20��40min, make DNA1 be attached to electrode surface;
E. step (d) gained glass-carbon electrode is immersed in the bovine serum albumin solution that 10��20 �� L mass percentage concentration are 2%, closes 1��2h (closing non-active site point), obtain the immobilized glass-carbon electrode having DNA1;
(2) assembling of solid-state electrochemistry illumination sensor
A. prepare a series of different concns containing the standardized solution of mercury ion, by 0.01��0.1MPBS damping fluid of standardized solution and pH=7��8 containing 10 ��m of ol/LDNA2 by volume 1:1 mix;
B. get mixing solutions 10��20 �� L of step (a) gained, it is added drop-wise to the immobilized glassy carbon electrode surface having DNA1, after incubated at room temperature 20��30min, by 0.01��0.1MPBS buffer solution for cleaning of pH=7��8;
C. get the avidin solution of 0.01��0.1MPBS damping fluid preparation 0.01��0.1mg/mL of appropriate pH=7��8,10��20 �� L avidin solution are added drop-wise to step (b) gained glassy carbon electrode surface, after hatching 5min;
D. after 0.01��0.1MPBS buffer solution for cleaning of glass-carbon electrode pH=7��8 of step (c) gained, the solid-state electrochemistry illumination sensor for mercury ion detecting will namely be obtained.
The structural formula of described DNA1 is: 5 '-NH2-(CH2)6-GACTGTCTCGTTCGCTTAG-3 '; The structural formula of described DNA2 is: 5 '-biotin-CTATGCGTACGTGACTGTC-3 '.
The method for detecting mercury ion of above-mentioned solid-state electrochemistry illumination sensor, concrete steps are as follows:
(1) typical curve is set up
A. after being rinsed well with 0.01��0.1MPBS damping fluid by the glass-carbon electrode of the immobilized DNA1 of having according to claim 3, in 0.05��0.2M carbonate buffer solution of pH=9��10, electrochemical reaction is started, test electrochemiluminescence intensity I0;
B. the solid-state electrochemistry illumination sensor for mercury ion detecting according to claim 3 is put into 0.05��0.2M carbonate buffer solution of pH=9��10, starts electrochemical reaction, measure electrochemiluminescence intensity, obtain the Hg of a series of different concns2+The electrochemiluminescence intensity level I that solution is corresponding1, calculate the Hg of a series of different concns2+The changes values �� I=I of solution electrochemistry luminous intensity0-I1, set up changes values �� I and the Hg of electrochemiluminescence intensity2+Quantitative relationship between strength of solution;
(2) testing sample measures
A. by containing mercury ion testing sample solution with contain 10 ��m of ol/LDNA2 pH7��8 0.01��0.1MPBS damping fluid by volume 1:1 mix;
B. get mixing solutions 10��20 �� L of step (a) gained, it is added drop-wise to the immobilized glassy carbon electrode surface having DNA1, after incubated at room temperature 20��30min, by 0.01��0.1MPBS buffer solution for cleaning of pH=7��8;
The avidin solution of the 0.01��0.1mg/mL c. prepared by 0.01��0.1MPBS damping fluid of 10��20 �� L pH=7��8 is added drop-wise to step (b) gained glassy carbon electrode surface, after hatching 5min;
D. by after 0.01��0.1MPBS buffer solution for cleaning of glass-carbon electrode pH=7��8 of step (c) gained, as working electrode, saturated calomel electrode or Ag/AgCl electrode are reference electrode, platinum wire electrode is to electrode, build three-electrode system, insert in 0.05��0.2M carbonate buffer solution of pH=9��10, start electrochemical reaction, test electrochemiluminescence intensity level, utilizes changes values �� I and the Hg of electrochemiluminescence intensity2+Quantitative relationship between strength of solution, calculates Hg in testing sample solution2+Actual concentrations CHg��
Containing the hydrogen peroxide of 1mM in described carbonate buffer solution, the electrochemical method of employing is potential step chronoamperometry; Potential step: 0V step is to 1V; Pulse width: 0.25s; Measuring Time interval: 30s.
Inventive principle: electrochemiluminescence reagent be the ABEI of electropolymerization, ABEI after glassy carbon electrode surface electropolymerization, two-step electrochemical luminous reaction originally turns into a step, and turns into reversible from irreversible, and electrochemiluminescence signal is highly stable. Amino in ABEI still retains, very convenient follow-up coupling DNA. Under glutaraldehyde cross-linking effect, the amino coupled in the amino of DNA1 and electrode surface ABEI polymkeric substance, thus it is fixed to electrode surface, the electrochemiluminescence intensity I now recorded0Very big. When solution to be measured has Hg2+Time, DNA1 and DNA2 passes through T-Hg2+-T mispairing combines, and avidin is combined with the vitamin H of DNA2 end, and vitamin H is biomacromolecule, and molecular weight is about 60kD, and it can effectively hinder the transmission of electronics and light at electrode surface, and electrochemiluminescence is weakened as I1. Hg solution to be measured2+Content is more high, the changes values �� I=I of luminous intensity0-I1Also more big, this is exactly this solid-state electrochemistry illumination sensor detection by quantitative Hg2+Mechanism. Solid-state electrochemistry illumination sensor detection Hg2+Schematic diagram as shown in Figure 1.
Compared with prior art, it is an advantage of the current invention that:
(1) highly sensitive, by means of the extremely high sensitivity of electrochemiluminescence technology itself, this solid-state electrochemistry illumination sensor energy detection by quantitative 0.01nMHg2+��
(2) highly selective, common metal ion is such as Pb2+��Mn2+��Co2+��Ni2+��Cu2+��Zn2+��Cd2+��Mg2+Detection is all noiseless. Reason is: T-Hg2+Mercury ion is had the recognition capability of high specific by-T mispairing, and the interference of other metal ion can be ignored.
(3) with low cost. Required amount of reagent is few.
(4) precision height. Adopting solid-state electrochemistry illumination, signal is stablized, result precision height.
In sum, the present invention draws up standby a kind of solid-state electrochemistry illumination sensor for mercury ion detecting, have highly sensitive, good stability, selectivity are strong, circulation ratio is good, be easy to the advantage of operation, it is possible to realization is to the testing goal of ultralow density mercury ion.
Accompanying drawing explanation
Fig. 1 is the schematic diagram of solid-state electrochemistry illumination sensor of the present invention detection mercury ion;
Fig. 2 is the electrochemiluminescence signal that different concns mercury ion is corresponding;
Fig. 3 is the linear relationship chart between the changes values �� I of luminous intensity and ion concentration of mercury logarithm;
Fig. 4 is the selective enumeration method result figure of solid-state electrochemistry illumination sensor of the present invention detection mercury ion.
Embodiment
Below in conjunction with accompanying drawing embodiment, the present invention is described in further detail.
Specific embodiment one
For detecting a solid-state electrochemistry illumination sensor for mercury ion, this sensor is the ABEI of surperficial immobilized electropolymerization, then be assembled with successively glutaraldehyde, DNA1, by T-Hg2+The vitamin H of DNA2, DNA2 end that-T mispairing combines and the glass-carbon electrode of avidin, concrete preparation process is as follows:
(1) the immobilized preparation having DNA1 glass-carbon electrode
A. by diameter be the glass-carbon electrode of 3��5mm successively with the aluminium sesquioxide polished finish of 1.0 ��m, 0.3 ��m, 0.05 ��m, then successively with ethanol, water ultrasonic cleaning, after water is rinsed well, nitrogen blows dry for subsequent use;
B. step (a) gained glass-carbon electrode is placed in the H containing N-(4-ammonia butyl) the different luminol,3-aminophthalic acid cyclic hydrazide of-N-ethyl (ABEI)2SO4In solution, carrying out cyclic voltammetry scan, voltage range is-0.2��1.5V, encloses with the speed scan round 20 of sweeping of 0.05V/s, makes ABEI at glassy carbon electrode surface electropolymerization; The above-mentioned H containing N-(4-ammonia butyl) the different luminol,3-aminophthalic acid cyclic hydrazide of-N-ethyl (ABEI)2SO4In solution, N-(4-ammonia butyl)-N-ethyl different luminol,3-aminophthalic acid cyclic hydrazide (ABEI) concentration is 0.001mol/L, H2SO4Concentration is 0.1��1mol/L;
C. after 0.01��0.1MPBS damping fluid of step (b) gained glass-carbon electrode pH=7��8 being rinsed well, drip in glassy carbon electrode surface and add 1��3wt% glutaraldehyde solution 10��20 �� L, leave standstill 20��40 minutes;
D. after 0.01��0.1MPBS damping fluid of step (c) gained glass-carbon electrode pH=7��8 being rinsed well, 0.01��0.1MPBS damping fluid 10��20 �� L of pH=7��8 added containing 10 ��m of ol/LDNA1 is dripped in glassy carbon electrode surface, leave standstill 20��40min, make DNA1 be attached to electrode surface; Wherein the structural formula of DNA1 is: 5 '-NH2-(CH2)6-GACTGTCTCGTTCGCTTAG-3 ';
E. step (d) gained glass-carbon electrode is immersed in the bovine serum albumin solution that 10��20 �� L mass percentage concentration are 2%, closes 1��2h (closing non-active site point), obtain the immobilized glass-carbon electrode having DNA1;
(2) assembling of solid-state electrochemistry illumination sensor
A. prepare a series of different concns containing the standardized solution of mercury ion, by 0.01��0.1MPBS damping fluid of standardized solution and pH=7��8 containing 10 ��m of ol/LDNA2 by volume 1:1 mix; Wherein the structural formula of DNA2 is: 5 '-biotin-CTATGCGTACGTGACTGTC-3 ';
B. get mixing solutions 10��20 �� L of step (a) gained, it is added drop-wise to the immobilized glassy carbon electrode surface having DNA1, after incubated at room temperature 20��30min, by 0.01��0.1MPBS buffer solution for cleaning of pH=7��8;
C. get the avidin solution of 0.01��0.1MPBS damping fluid preparation 0.01��0.1mg/mL of appropriate pH=7��8,10��20 �� L avidin solution are added drop-wise to step (b) gained glassy carbon electrode surface, after hatching 5min;
D. after 0.01��0.1MPBS buffer solution for cleaning of glass-carbon electrode pH=7��8 of step (c) gained, the solid-state electrochemistry illumination sensor for mercury ion detecting will namely be obtained.
Except above-mentioned specific embodiment, in step prepared by above-mentioned solid-state electrochemistry illumination sensor:
The speed of sweeping of cyclic voltammetry scan can also be any value in 0.01��0.1V/s, and the number of turns can also be any value in 20��30 circles, containing the H of N-(4-ammonia butyl) the different luminol,3-aminophthalic acid cyclic hydrazide of-N-ethyl (ABEI)2SO4In solution, N-(4-ammonia butyl)-N-ethyl different luminol,3-aminophthalic acid cyclic hydrazide (ABEI) concentration can also be any value in 0.01��0.0001mol/L, H2SO4Concentration can be any value in 0.1��1mol/L;
The concentration of glutaraldehyde solution can be any value in 1��3wt%, drips that to add volume can be any value in 10��20 �� L, and time of repose is any value in 20��40 minutes;
Containing 10 ��m of ol/LDNA1 pH=7��8 0.01��0.1MPBS damping fluid drip that to add volume can be any value in 10��20 �� L, time of repose can be any value in 20��40 minutes;
Bovine serum albumin (BSA) solution drips that to add volume can be any value in 10��20 �� L, and time of repose can be any value in 1��2h;
The dripping of glassy carbon electrode surface that standardized solution is added drop-wise to the immobilized DNA1 of having with 0.01��0.1MPBS damping fluid gained mixing solutions of 1:1 mixing by volume of pH=7��8 containing 10 ��m of ol/LDNA2 adds any value that volume can be 10��20 �� L, and incubation time can be any value in 20��30min;
The concentration of avidin solution can be any value in 0.01��0.1mg/mL, drips that to add volume can be any value in 10��20 �� L;
The concentration of PBS can be any value in 0.01��0.1M, and pH can be any value in 7��8.
Specific embodiment two
The solid-state electrochemistry illumination sensor utilizing above-mentioned specific embodiment two to prepare detects the method for mercury ion, and as shown in Figure 1, concrete steps are as follows for the principle of detection mercury ion:
After glass-carbon electrode 0.01��0.1MPBS damping fluid of the immobilized DNA1 of having a. prepared by above-mentioned specific embodiment two is rinsed well, in 0.05��0.2M carbonate buffer solution of pH=9��10, start electrochemical reaction, test electrochemiluminescence intensity I0;
B. the solid-state electrochemistry illumination sensor being used for mercury ion detecting that above-mentioned specific embodiment two prepares is put into 0.05��0.2M carbonate buffer solution of pH=9��10, start electrochemical reaction, measure electrochemiluminescence intensity, obtain the Hg of a series of different concns2+The electrochemiluminescence intensity level I that solution is corresponding1, calculate the Hg of a series of different concns2+The changes values �� I=I of solution electrochemistry luminous intensity0-I1,
The changes values �� I=I of electrochemiluminescence intensity0-I1It is, linear in 0.01��10nM concentration range with ion concentration of mercury logarithm that (the electrochemiluminescence signal that different concns mercury ion is corresponding, as shown in Figure 2), typical curve is �� I=2493.25+843.63logCHg(nM) (linear relationship chart between the changes values �� I of luminous intensity and ion concentration of mercury logarithm, as shown in Figure 3);
(2) testing sample measures
A. by containing mercury ion testing sample solution with contain 10 ��m of ol/LDNA2 pH7��8 0.01��0.1MPBS damping fluid by volume 1:1 mix;
B. mixing solutions 10��20 �� L of step (a) gained is got, it is added drop-wise to the immobilized glassy carbon electrode surface having DNA1 that above-mentioned specific embodiment one prepares, after incubated at room temperature 20��30min, by 0.01��0.1MPBS buffer solution for cleaning of pH=7��8;
The avidin solution of the 0.01��0.1mg/mL c. prepared by 0.01��0.1MPBS damping fluid of 10��20 �� L pH=7��8 is added drop-wise to step (b) gained glassy carbon electrode surface, after hatching 5min;
D. by after 0.01��0.1MPBS buffer solution for cleaning of glass-carbon electrode pH=7��8 of step (c) gained, as working electrode, saturated calomel electrode or Ag/AgCl electrode are reference electrode, platinum wire electrode is to electrode, build three-electrode system, insert in 0.05��0.2M carbonate buffer solution of pH=9��10, start electrochemical reaction, test electrochemiluminescence intensity level, according to typical curve �� I=2493.25+843.63logCHg(nM) Hg in testing sample solution, is calculated2+Actual concentrations CHg, unit is nM.
Containing the hydrogen peroxide of 1mM in above-mentioned carbonate buffer solution, the electrochemical method of employing is potential step chronoamperometry; Potential step: 0V step is to 1V; Pulse width: 0.25s; Measuring Time interval: 30s.
Specific embodiment three
The detection experiment of highly selective and high sensitivity
High sensitivity is embodied by specific embodiment two, due to the changes values �� I=I of electrochemiluminescence intensity0-I1, with ion concentration of mercury logarithm in 0.01��10nM concentration range linear, therefore, the solid-state electrochemistry illumination sensor of the detection mercury ion that specific embodiment one prepares is to Hg2+Detection sensitivity reach 0.01nM.
Highly selective: with the experiment condition that above-mentioned specific embodiment two is same, detects the common interference ion of 10 ��Ms: Pb2+,Mn2+,Co2+,Ni2+,Cu2+,Zn2+,Cd2+,Mg2+, result is as shown in Figure 4.
Result shows: 0.01nMHg2+The �� I causing sensor is about 800, and the �� I that the common interference ion of 10 ��Ms causes sensor is all less than 200, it is meant that the common interference ion of 1000000 times does not affect detection, and major cause is Hg2+And the T-Hg between T base2+Specific recognition between-T mispairing, vitamin H and avidin.
Specific embodiment four
The solid-state electrochemistry illumination sensor of the detection mercury ion prepared taking above-mentioned specific embodiment one, as working electrode, with the experiment condition that above-mentioned specific embodiment two is same, detects the Hg of high, normal, basic three kinds of concentration2+Standardized solution, result is such as following table.
As seen from the above table, the rate of recovery, between 90.8��97.4%, shows that accuracy is good. RSD, between 6.4��11.0%, shows that precision is good.
Above-mentioned explanation is limitation of the present invention not, and the present invention is also not limited to above-mentioned citing. Those skilled in the art are in the essential scope of the present invention, and change, remodeling, interpolation or the replacement made, also should belong to protection scope of the present invention, and protection scope of the present invention is as the criterion with claim book.
Claims (6)
1. one kind for detecting the solid-state electrochemistry illumination sensor of mercury ion, it is characterized in that: immobilized N-(4-ammonia butyl) the different luminol,3-aminophthalic acid cyclic hydrazide of-N-ethyl (ABEI) having electropolymerization of working electrode surface, described N-(4-ammonia butyl) the different luminol,3-aminophthalic acid cyclic hydrazide of-N-ethyl is assembled with successively glutaraldehyde, DNA1, by T-Hg2+The vitamin H of DNA2, DNA2 end that-T mispairing combines and avidin.
2. the solid-state electrochemistry illumination sensor for detecting mercury ion according to claim 1, it is characterised in that: described working electrode is glass-carbon electrode.
3. the preparation method of the solid-state electrochemistry illumination sensor for detecting mercury ion according to claim 1, it is characterised in that concrete steps are as follows:
(1) the immobilized preparation having DNA1 glass-carbon electrode
A. by diameter be the glass-carbon electrode of 3��5mm successively with the aluminium sesquioxide polished finish of 1.0 ��m, 0.3 ��m, 0.05 ��m, then successively with ethanol, water ultrasonic cleaning, after water is rinsed well, nitrogen blows dry for subsequent use;
B. step (a) gained glass-carbon electrode is placed in the H containing N-(4-ammonia butyl) the different luminol,3-aminophthalic acid cyclic hydrazide of-N-ethyl (ABEI)2SO4In solution, carrying out cyclic voltammetry scan, voltage range is-0.2��1.5V, encloses with the speed scan round 20��30 of sweeping of 0.01��0.1V/s, makes ABEI at glassy carbon electrode surface electropolymerization; The described H containing N-(4-ammonia butyl) the different luminol,3-aminophthalic acid cyclic hydrazide of-N-ethyl (ABEI)2SO4In solution, N-(4-ammonia butyl)-N-ethyl different luminol,3-aminophthalic acid cyclic hydrazide (ABEI) concentration is 0.01��0.0001mol/L, H2SO4Concentration is 0.1��1mol/L;
C. after 0.01��0.1MPBS damping fluid of step (b) gained glass-carbon electrode pH=7��8 being rinsed well, drip in glassy carbon electrode surface and add 1��3wt% glutaraldehyde solution 10��20 �� L, leave standstill 20��40 minutes;
D. after 0.01��0.1MPBS damping fluid of step (c) gained glass-carbon electrode pH=7��8 being rinsed well, 0.01��0.1MPBS damping fluid 10��20 �� L of pH=7��8 added containing 10 ��m of ol/LDNA1 is dripped in glassy carbon electrode surface, leave standstill 20��40min, make DNA1 be attached to electrode surface;
E. step (d) gained glass-carbon electrode is immersed in the bovine serum albumin solution that 10��20 �� L mass percentage concentration are 2%, closes 1��2h, obtain the immobilized glass-carbon electrode having DNA1;
(2) assembling of solid-state electrochemistry illumination sensor
A. prepare a series of different concns containing the standardized solution of mercury ion, by 0.01��0.1MPBS damping fluid of standardized solution and pH=7��8 containing 10 ��m of ol/LDNA2 by volume 1:1 mix;
B. get mixing solutions 10��20 �� L of step (a) gained, it is added drop-wise to the immobilized glassy carbon electrode surface having DNA1, after incubated at room temperature 20��30min, by 0.01��0.1MPBS buffer solution for cleaning of pH=7��8;
C. get the avidin solution of 0.01��0.1MPBS damping fluid preparation 0.01��0.1mg/mL of appropriate pH=7��8,10��20 �� L avidin solution are added drop-wise to step (b) gained glassy carbon electrode surface, after hatching 5min;
D. after 0.01��0.1MPBS buffer solution for cleaning of glass-carbon electrode pH=7��8 of step (c) gained, the solid-state electrochemistry illumination sensor for mercury ion detecting will namely be obtained.
4. the preparation method of a kind of solid-state electrochemistry illumination sensor for detecting mercury ion according to claim 3, it is characterised in that: the structural formula of described DNA1 is: 5 '-NH2-(CH2)6-GACTGTCTCGTTCGCTTAG-3 '; The structural formula of described DNA2 is: 5 '-biotin-CTATGCGTACGTGACTGTC-3 '.
5. the detection method of the solid-state electrochemistry illumination sensor of a detection mercury ion according to claim 3 or 4, it is characterised in that concrete steps are as follows:
(1) typical curve is set up
A. after being rinsed well with 0.01��0.1MPBS damping fluid by the glass-carbon electrode of the immobilized DNA1 of having according to claim 3, in 0.05��0.2M carbonate buffer solution of pH=9��10, electrochemical reaction is started, test electrochemiluminescence intensity I0;
B. the solid-state electrochemistry illumination sensor for mercury ion detecting according to claim 3 is put into 0.05��0.2M carbonate buffer solution of pH=9��10, starts electrochemical reaction, measure electrochemiluminescence intensity, obtain the Hg of a series of different concns2+The electrochemiluminescence intensity level I that solution is corresponding1, calculate the Hg of a series of different concns2+The changes values �� I=I of solution electrochemistry luminous intensity0-I1, set up changes values �� I and the Hg of electrochemiluminescence intensity2+Quantitative relationship between strength of solution;
(2) testing sample measures
A. by containing mercury ion testing sample solution with contain 10 ��m of ol/LDNA2 pH7��8 0.01��0.1MPBS damping fluid by volume 1:1 mix;
B. get mixing solutions 10��20 �� L of step (a) gained, it is added drop-wise to the immobilized glassy carbon electrode surface having DNA1, after incubated at room temperature 20��30min, by 0.01��0.1MPBS buffer solution for cleaning of pH=7��8;
The avidin solution of the 0.01��0.1mg/mL c. prepared by 0.01��0.1MPBS damping fluid of 10��20 �� L pH=7��8 is added drop-wise to step (b) gained glassy carbon electrode surface, after hatching 5min;
D. by after 0.01��0.1MPBS buffer solution for cleaning of glass-carbon electrode pH=7��8 of step (c) gained, as working electrode, saturated calomel electrode or Ag/AgCl electrode are reference electrode, platinum wire electrode is to electrode, build three-electrode system, insert in 0.05��0.2M carbonate buffer solution of pH=9��10, start electrochemical reaction, test electrochemiluminescence intensity level, according to changes values �� I and the Hg of electrochemiluminescence intensity2+Quantitative relationship between strength of solution, calculates Hg in testing sample solution2+Actual concentrations CHg��
6. the detection method of a kind of solid-state electrochemistry illumination sensor detecting mercury ion according to claim 5, it is characterised in that: containing the hydrogen peroxide of 1mM in described carbonate buffer solution, the electrochemical method of employing is potential step chronoamperometry; Potential step: 0V step is to 1V; Pulse width: 0.25s; Measuring Time interval: 30s.
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| CN106770590A (en) * | 2016-11-17 | 2017-05-31 | 江苏智石科技有限公司 | A kind of Mercury in Soil concentration of metal ions monitoring system based on three-stage amplifier |
| CN107543852B (en) * | 2017-08-29 | 2019-11-29 | 济南大学 | A kind of Electrochemiluminescsensor sensor based on functional metal organic framework materials |
| CN109470751B (en) * | 2018-09-13 | 2020-10-30 | 宁波大学 | Preparation method and application of electrochemical sensor for detecting mercury ions through dual-channel output |
| CN110006968B (en) * | 2019-03-12 | 2021-03-09 | 宁波大学 | Preparation method and application of electrochemical biosensor for detecting mercury ions based on rapid scanning cyclic voltammetry technology |
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