CN104155289A - Solid electrochemical luminescence sensor for detecting mercury ions and preparation method and application of solid electrochemical luminescence sensor for detecting mercury ions - Google Patents

Solid electrochemical luminescence sensor for detecting mercury ions and preparation method and application of solid electrochemical luminescence sensor for detecting mercury ions Download PDF

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CN104155289A
CN104155289A CN201410289966.1A CN201410289966A CN104155289A CN 104155289 A CN104155289 A CN 104155289A CN 201410289966 A CN201410289966 A CN 201410289966A CN 104155289 A CN104155289 A CN 104155289A
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glass
solution
carbon electrode
solid
dna1
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CN104155289B (en
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郭智勇
陈贝贝
沙玉红
徐成银
郝婷婷
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Leling Jinshan Electric Appliance Co ltd
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Ningbo University
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Abstract

The invention discloses a solid electrochemical luminescence sensor for detecting mercury ions and a preparation method and application of the solid electrochemical luminescence sensor for detecting the mercury ions. The solid electrochemical luminescence sensor is characterized by being a glassy carbon electrode, wherein the surface of the glassy carbon electrode is immobilized with electrically polymerized ABEI, and then the glassy carbon electrode is assembled with glutaraldehyde, DNA1, T-Hg<2+>-T mismatched DNA2 and biotin. The preparation method comprises the steps: preparing the glassy carbon electrode immobilized with the DNA1 and assembling the solid electrochemical luminescence sensor. The solid electrochemical luminescence sensor has the advantages of high sensitivity, high stability, high selectivity, high repetitiveness, easiness in operation and the like, and reagents can be saved.

Description

A kind of solid-state electrochemistry illumination sensor that detects mercury ion and its preparation method and application
Technical field
The present invention relates to heavy metal detection technique field, especially relate to a kind of solid-state electrochemistry illumination sensor that detects mercury ion and its preparation method and application.
Background technology
Heavy metal pollution is one of important component part of present environmental pollution.Mercury (Hg), is liquid metal unique under normal temperature, is the No.1 killer in heavy metal pollution.In recent years, the mankind are increasing to the exploitation of heavy metal Hg, smelting, processing and business manufacturing activities, and a large amount of mercury enters in atmosphere, 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 biosome is caused to very major injury, produces neuropsychic symptom, trembles, the disease such as stomatitis and toxic nephropathy.Mercury ion (Hg 2+) be mercury in one of main existence form of occurring in nature, in numerous hygienic standards, requirement must not detect, and the low concentration mercury ion detecting method that therefore develops high sensitivity and high selectivity has great importance to environment measuring and food safety monitoring.
The method that detects at present mercury ion mainly contains: cold atomic absorption spectrometry, ICP-MS, fluorescent spectrometry, ultraviolet-visible spectrophotometry, atomic fluorescence method, electrochemical process, the chromatography of ions, capillary electrophoresis, heavy metal rapid detector method, test strips method etc., but these methods maybe need to use expensive instrument, complex operation, and require testing staff to possess certain professional knowledge, analysis cost is high, is difficult to popularize; Or sensitivity is not high, poor selectivity, sample pretreatment is complicated, is difficult to accurate quantitative analysis and detects low concentration Hg 2+.
Electrochemiluminescence (ECL) is a kind of analytical technology that galvanochemistry is combined with chemiluminescence, highly sensitive, the advantage such as the range of linearity is wide, observation is convenient, instrument is simple not only with chemiluminometry, and the current potential controllability with electrochemical methods is strong, selectivity is high, stable reagent, high repeatability and other advantages, receive the very big concern of researcher, and developed into gradually the important branch in analyzing and testing field.Solid-state electrochemistry illumination is by chemical method or physical method is immobilized carries out the technology of ECL detection after electrode surface again by electrochemiluminescence reagent, compare with the ECL in solution, electrochemiluminescence reagent is fixed to the solid-state ECL that electrode surface builds, also has following advantage: the consumption, simplification experimental implementation, the ECL intensity etc. that improves that reduce expensive reagent.But at present, the solid-state electrochemistry illumination that does not also disclose any N-of utilization (4-ammonia butyl)-different luminol of N-ethyl (ABEI) both at home and abroad detects Mercury in Water Body ion concentration relevant report.
Summary of the invention
Technical matters to be solved by this invention is to provide that a kind of highly sensitive, good stability, selectivity are strong, the solid-state electrochemistry illumination sensor for mercury ion detecting of favorable reproducibility, easy operating and preparation method thereof and detection method.
The present invention solves the problems of the technologies described above adopted technical scheme: a kind of solid-state electrochemistry illumination sensor for detection of mercury ion, and the ABEI that described sensor is surperficial immobilized electropolymerization, then be assembled with successively glutaraldehyde, DNA1, pass through T-Hg 2+the DNA2 of-T mispairing combination and the glass-carbon electrode of biotin.
The preparation method of the above-mentioned solid-state electrochemistry illumination sensor for detection of mercury ion, concrete steps are as follows:
(1) the immobilized preparation that has DNA1 glass-carbon electrode
A. by diameter, be the alundum (Al2O3) polishing that the glass-carbon electrode of 3～5 mm is used 1.0 μ m, 0.3 μ m, 0.05 μ m successively, then use successively ethanol, water ultrasonic cleaning, after water is rinsed well, nitrogen dries up standby;
B. step (a) gained glass-carbon electrode is placed in to the H containing N-(4-ammonia butyl)-different luminol of N-ethyl (ABEI) 2sO 4in solution, carry out cyclic voltammetry scan, voltage range is 0.2～1.5 V, with fast scan round 20～30 circles of sweeping of 0.01～0.1 V/s, makes ABEI in the polymerization of glass-carbon electrode surface electrical; The described H containing N-(4-ammonia butyl)-different luminol of N-ethyl (ABEI) 2sO 4n-in solution (4-ammonia the butyl)-different luminol of N-ethyl (ABEI) concentration is 0.01～0.0001 mol/L, H 2sO 4concentration is 0.1～1 mol/L;
C., after 0.01～0.1 M PBS damping fluid of pH=7～8 is rinsed well for glass-carbon electrode by step (b) gained, on glass-carbon electrode surface, drip 1～3 wt% glutaraldehyde solution 10～20 μ L, standing 20～40 minutes;
D. after by step (c) gained, for glass-carbon electrode, 0.01～0.1 M PBS damping fluid of pH=7～8 is rinsed well, on glass-carbon electrode surface, drip 0.01～0.1 M PBS damping fluid 10～20 μ L containing pH=7～8 of 10 μ mol/L DNA1, standing 20～40 min, 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 μ L mass percentage concentration are 2%, seals 1～2 h (sealing nonactive site), obtain the glass-carbon electrode of the immobilized DNA1 of having;
(2) assembling of solid-state electrochemistry illumination sensor
A. prepare a series of variable concentrations containing the standard solution of mercury ion, by standard solution with contain 10 μ mol/L DNA2 pH=7～8 0.01～0.1 M PBS damping fluid by volume 1:1 mix;
B. get mixed solution 10～20 μ L of step (a) gained, be added drop-wise to the glass-carbon electrode surface of the immobilized DNA1 of having, under room temperature, hatch after 20～30 min, by 0.01～0.1 M PBS buffer solution for cleaning of pH=7～8;
C. the Avidin solution of getting 0.01～0.1 M PBS damping fluid preparation 0.01～0.1 mg/mL of appropriate pH=7～8, is added drop-wise to step (b) gained glass-carbon electrode surface by 10～20 μ L Avidin solution, hatches after 5 min;
D. by after 0.01～0.1 M PBS buffer solution for cleaning of pH=7～8 for glass-carbon electrode of step (c) gained, obtain the solid-state electrochemistry illumination sensor for mercury ion detecting.
The structural formula of described DNA1 is: 5 '-NH 2-(CH 2) 6-GAC tgTC tcGT tcGC ttAG-3 '; The structural formula of described DNA2 is: 5 '-biotin-CTA tgCG taCG tgAC tgTC-3 '.
The method for detection of mercury ion of above-mentioned solid-state electrochemistry illumination sensor, concrete steps are as follows:
(1) typical curve is set up
A. after the glass-carbon electrode of the immobilized DNA1 of having claimed in claim 3 being rinsed well with 0.01～0.1 M PBS damping fluid, in 0.05～0.2 M carbonate buffer solution of pH=9～10, start electrochemical reaction, test electrochemiluminescence intensity I 0;
B. the solid-state electrochemistry illumination sensor for mercury ion detecting claimed in claim 3 is put into 0.05～0.2 M carbonate buffer solution of pH=9～10, started electrochemical reaction, measure electrochemiluminescence intensity, obtain the Hg of a series of variable concentrations 2+the electrochemiluminescence intensity level I that solution is corresponding 1, calculate the Hg of a series of variable concentrations 2+the change value Δ I=I of solution electrochemistry luminous intensity 0i 1, set up change value Δ I and the Hg of electrochemiluminescence intensity 2+quantitative relationship between solution concentration;
(2) testing sample is measured
A. by containing the testing sample solution of mercury ion with contain 10 μ mol/L DNA2 pH7～8 0.01～0.1 M PBS damping fluid by volume 1:1 mix;
B. get mixed solution 10～20 μ L of step (a) gained, be added drop-wise to the glass-carbon electrode surface of the immobilized DNA1 of having, under room temperature, hatch after 20～30 min, by 0.01～0.1 M PBS buffer solution for cleaning of pH=7～8;
C. the Avidin solution of 10～20 μ, 0.01～0.1 mg/mL of 0.01～0.1 M PBS damping fluid preparation of pH=7～8 for L is added drop-wise to step (b) gained glass-carbon electrode surface, hatches after 5 min;
D. by after 0.01～0.1 M PBS buffer solution for cleaning of pH=7～8 for glass-carbon electrode of step (c) gained, as working electrode, saturated calomel electrode or Ag/AgCl electrode are contrast electrode, platinum electrode is to electrode, build three-electrode system, insert in 0.05～0.2 M carbonate buffer solution of pH=9～10, start electrochemical reaction, test electrochemiluminescence intensity level, utilize change value Δ I and the Hg of electrochemiluminescence intensity 2+quantitative relationship between solution concentration, calculates Hg in testing sample solution 2+accurate concentration C hg.
In described carbonate buffer solution, containing the hydrogen peroxide of 1 mM, the electrochemical method of employing is potential step chronoamperometry; Potential step: 0 V step to 1 V; Pulse width: 0.25 s; Measuring intervals of TIME: 30 s.
Inventive principle: the ABEI that electrochemiluminescence reagent is electropolymerization, ABEI is after the polymerization of glass-carbon electrode surface electrical, and two-step electrochemical luminescence-producing reaction originally becomes a step, and never reversiblely becomes reversiblely, and electrochemical luminescence signals 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 be fixed to electrode surface, the electrochemiluminescence intensity I now recording 0very large.In solution to be measured, there is Hg 2+time, DNA1 and DNA2 pass through T-Hg 2+-T mispairing combination, Avidin is combined with the biotin of DNA2 end, and biotin is biomacromolecule, the about 60kD of molecular weight, it can effectively hinder the transmission of electronics and light at electrode surface, and electrochemiluminescence is weakened as I 1.Hg solution to be measured 2+content is higher, the change value Δ I=I of luminous intensity 0i 1also just larger, Here it is, and this solid-state electrochemistry illumination sensor quantitatively detects Hg 2+mechanism.Solid-state electrochemistry illumination sensor detects Hg 2+schematic diagram as shown in Figure 1.
Compared with prior art, the invention has the advantages that:
(1) high sensitivity, by means of the high sensitivity of electrochemiluminescence technology itself, this solid-state electrochemistry illumination sensor can quantitatively detect 0.01 nM Hg 2+.
(2) high selectivity, common metal ion is as Pb 2+, Mn 2+, Co 2+, Ni 2+, Cu 2+, Zn 2+, Cd 2+, Mg 2+all noiseless to detecting.Reason is: T-Hg 2+-T mispairing has the recognition capability of high specific to mercury ion, the interference of other metallic ion can be ignored.
(3) with low cost.Required amount of reagent is few.
(4) precision is high.Adopt solid-state electrochemistry illumination, signal stabilization, result precision is high.
In sum, the present invention's standby a kind of solid-state electrochemistry illumination sensor for mercury ion detecting of drawing up, has advantages of highly sensitive, good stability, selectivity is strong, reappearance is good, easy operating, can realize the testing goal to super low concentration mercury ion.
Accompanying drawing explanation
Fig. 1 is the schematic diagram that solid-state electrochemistry illumination sensor of the present invention detects mercury ion;
Fig. 2 is the electrochemical luminescence signals that variable concentrations mercury ion is corresponding;
Fig. 3 is the change value Δ I of luminous intensity and the linear relationship chart between ion concentration of mercury logarithm;
Fig. 4 is the selectivity testing result figure that solid-state electrochemistry illumination sensor of the present invention detects mercury ion.
Embodiment
Below in conjunction with accompanying drawing, embodiment is described in further detail the present invention.
Specific embodiment one
For detection of a solid-state electrochemistry illumination sensor for mercury ion, the ABEI that this sensor is surperficial immobilized electropolymerization, then be assembled with successively glutaraldehyde, DNA1, pass through T-Hg 2+the DNA2 of-T mispairing combination and the glass-carbon electrode of biotin, concrete preparation process is as follows:
(1) the immobilized preparation that has DNA1 glass-carbon electrode
A. by diameter, be the alundum (Al2O3) polishing that the glass-carbon electrode of 3～5 mm is used 1.0 μ m, 0.3 μ m, 0.05 μ m successively, then use successively ethanol, water ultrasonic cleaning, after water is rinsed well, nitrogen dries up standby;
B. step (a) gained glass-carbon electrode is placed in to the H containing N-(4-ammonia butyl)-different luminol of N-ethyl (ABEI) 2sO 4in solution, carry out cyclic voltammetry scan, voltage range is 0.2～1.5 V, with fast scan round 20 circles of sweeping of 0.05 V/s, makes ABEI in the polymerization of glass-carbon electrode surface electrical; The above-mentioned H containing N-(4-ammonia butyl)-different luminol of N-ethyl (ABEI) 2sO 4n-in solution (4-ammonia the butyl)-different luminol of N-ethyl (ABEI) concentration is 0.001 mol/L, H 2sO 4concentration is 0.1 ~ 1 mol/L;
C., after 0.01～0.1 M PBS damping fluid of pH=7～8 is rinsed well for glass-carbon electrode by step (b) gained, on glass-carbon electrode surface, drip 1～3 wt% glutaraldehyde solution 10～20 μ L, standing 20～40 minutes;
D. after by step (c) gained, for glass-carbon electrode, 0.01～0.1 M PBS damping fluid of pH=7～8 is rinsed well, on glass-carbon electrode surface, drip 0.01～0.1 M PBS damping fluid 10～20 μ L containing pH=7～8 of 10 μ mol/L DNA1, standing 20～40 min, make DNA1 be attached to electrode surface; Wherein the structural formula of DNA1 is: 5 '-NH 2-(CH 2) 6-GAC tgTC tcGT tcGC ttAG-3 ';
E. step (d) gained glass-carbon electrode is immersed in the bovine serum albumin solution that 10～20 μ L mass percentage concentration are 2%, seals 1～2 h (sealing nonactive site), obtain the glass-carbon electrode of the immobilized DNA1 of having;
(2) assembling of solid-state electrochemistry illumination sensor
A. prepare a series of variable concentrations containing the standard solution of mercury ion, by standard solution with contain 10 μ mol/L DNA2 pH=7～8 0.01～0.1 M PBS damping fluid by volume 1:1 mix; Wherein the structural formula of DNA2 is: 5 '-biotin-CTA tgCG taCG tgAC tgTC-3 ';
B. get mixed solution 10～20 μ L of step (a) gained, be added drop-wise to the glass-carbon electrode surface of the immobilized DNA1 of having, under room temperature, hatch after 20～30 min, by 0.01～0.1 M PBS buffer solution for cleaning of pH=7～8;
C. the Avidin solution of getting 0.01～0.1 M PBS damping fluid preparation 0.01～0.1 mg/mL of appropriate pH=7～8, is added drop-wise to step (b) gained glass-carbon electrode surface by 10～20 μ L Avidin solution, hatches after 5 min;
D. by after 0.01～0.1 M PBS buffer solution for cleaning of pH=7～8 for glass-carbon electrode of step (c) gained, obtain the solid-state electrochemistry illumination sensor for mercury ion detecting.
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 the arbitrary value in 0.01～0.1 V/s, and the number of turns can also be for the arbitrary value in 20～30 circles, containing the H of N-(4-ammonia butyl)-different luminol of N-ethyl (ABEI) 2sO 4n-in solution (4-ammonia the butyl)-different luminol of N-ethyl (ABEI) concentration can also be the arbitrary value in 0.01 ~ 0.0001 mol/L, H 2sO 4concentration can be the arbitrary value in 0.1 ~ 1 mol/L;
The concentration of glutaraldehyde solution can be the arbitrary value in 1～3 wt%, and dripping volume can be the arbitrary value in 10～20 μ L, and time of repose is the arbitrary value in 20～40 minutes;
Dropping volume containing 0.01～0.1 M PBS damping fluid of pH=7～8 of 10 μ mol/L DNA1 can be the arbitrary value in 10～20 μ L, and time of repose can be the arbitrary value in 20～40 minutes;
It can be the arbitrary value in 10～20 μ L that bovine serum albumin(BSA) (BSA) solution drips volume, and time of repose can be the arbitrary value in 1～2 h;
Standard solution can be arbitrary value of 10～20 μ L with the dropping volume that 0.01～0.1 M PBS damping fluid 1:1 mixing by volume gained mixed solution that contains pH=7～8 of 10 μ mol/L DNA2 is added drop-wise to the glass-carbon electrode surface of the immobilized DNA1 of having, and incubation time can be the arbitrary value in 20～30 min;
The concentration of Avidin solution can be the arbitrary value in 0.01～0.1 mg/mL, and dripping volume can be the arbitrary value in 10～20 μ L;
The concentration of PBS damping fluid can be the arbitrary value in 0.01～0.1 M, and pH can be the arbitrary value in 7～8.
Specific embodiment two
Utilize solid-state electrochemistry illumination sensor that above-mentioned specific embodiment two prepares to detect the method for mercury ion, detect the principle of mercury ion as shown in Figure 1, concrete steps are as follows:
A. after the glass-carbon electrode of the immobilized DNA1 of having above-mentioned specific embodiment two being prepared is rinsed well with 0.01～0.1 M PBS damping fluid, in 0.05～0.2 M carbonate buffer solution of pH=9～10, start electrochemical reaction, test electrochemiluminescence intensity I 0;
B. the solid-state electrochemistry illumination sensor for mercury ion detecting above-mentioned specific embodiment two being prepared is put into 0.05～0.2 M carbonate buffer solution of pH=9～10, start electrochemical reaction, measure electrochemiluminescence intensity, obtain the Hg of a series of variable concentrations 2+the electrochemiluminescence intensity level I that solution is corresponding 1, calculate the Hg of a series of variable concentrations 2+the change value Δ I=I of solution electrochemistry luminous intensity 0i 1,
The change value Δ I=I of electrochemiluminescence intensity 0i 1it is, linear in 0.01～10 nM concentration range with ion concentration of mercury logarithm that (electrochemical luminescence signals that variable concentrations mercury ion is corresponding, as shown in Figure 2), typical curve is Δ I=2493.25+843.63 log C hg(nM) (the change value Δ I of luminous intensity and the linear relationship chart between ion concentration of mercury logarithm, as shown in Figure 3);
(2) testing sample is measured
A. by containing the testing sample solution of mercury ion with contain 10 μ mol/L DNA2 pH7～8 0.01～0.1 M PBS damping fluid by volume 1:1 mix;
B. get mixed solution 10～20 μ L of step (a) gained, be added drop-wise to the glass-carbon electrode surface of the immobilized DNA1 of having that above-mentioned specific embodiment one prepares, under room temperature, hatch after 20～30 min, by 0.01～0.1 M PBS buffer solution for cleaning of pH=7～8;
C. the Avidin solution of 10～20 μ, 0.01～0.1 mg/mL of 0.01～0.1 M PBS damping fluid preparation of pH=7～8 for L is added drop-wise to step (b) gained glass-carbon electrode surface, hatches after 5 min;
D. by after 0.01～0.1 M PBS buffer solution for cleaning of pH=7～8 for glass-carbon electrode of step (c) gained, as working electrode, saturated calomel electrode or Ag/AgCl electrode are contrast electrode, platinum electrode is to electrode, build three-electrode system, insert in 0.05～0.2 M carbonate buffer solution of pH=9～10, start electrochemical reaction, test electrochemiluminescence intensity level, according to typical curve Δ I=2493.25+843.63 log C hg(nM), calculate Hg in testing sample solution 2+accurate concentration C hg, unit is nM.
In above-mentioned carbonate buffer solution, containing the hydrogen peroxide of 1 mM, the electrochemical method of employing is potential step chronoamperometry; Potential step: 0 V step to 1 V; Pulse width: 0.25 s; Measuring intervals of TIME: 30 s.
Specific embodiment three
The detection test of high selectivity and high sensitivity
High sensitivity is embodied by specific embodiment two, due to the change value Δ I=I of electrochemiluminescence intensity 0i 1, linear in 0.01～10 nM concentration range with ion concentration of mercury logarithm, therefore, the solid-state electrochemistry illumination sensor of the detection mercury ion that specific embodiment one prepares is to Hg 2+detection sensitivity reach 0.01 nM.
High selectivity: with the same experiment condition of above-mentioned specific embodiment two, detect the common interference ion of 10 μ M: Pb 2+, Mn 2+, Co 2+, Ni 2+, Cu 2+, Zn 2+, Cd 2+, Mg 2+, result as shown in Figure 4.
Result shows: 0.01 nM Hg 2+causing the Δ I of sensor is 800 left and right, and the common interference ion of 10 μ M causes the Δ I of sensor to be all less than 200, means that the common interference ion of 1000000 times does not affect detection, and main cause is Hg 2+and the T-Hg between T base 2+specific recognition between-T mispairing, biotin and Avidin.
Specific embodiment four
The solid-state electrochemistry illumination sensor of the detection mercury ion that the above-mentioned specific embodiment one of take prepares is working electrode, with the same experiment condition of above-mentioned specific embodiment two, detects the Hg of high, normal, basic three kinds of concentration 2+standard solution, result is as following table.
As seen from the above table, the 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 not limitation of the present invention, and the present invention is also not limited to above-mentioned giving an example.Those skilled in the art are in essential scope of the present invention, and the variation of making, remodeling, interpolation or replacement, also should belong to protection scope of the present invention, and protection scope of the present invention is as the criterion with claims.

Claims (6)

1. the solid-state electrochemistry illumination sensor for detection of mercury ion, it is characterized in that: immobilized N-(4-ammonia the butyl)-different luminol of N-ethyl (ABEI) that has electropolymerization of described working electrode surface, is assembled with successively glutaraldehyde, DNA1 on described N-(4-ammonia butyl)-different luminol of N-ethyl, passes through T-Hg 2+dNA2 and the biotin of-T mispairing combination.
2. the solid-state electrochemistry illumination sensor for detection of mercury ion according to claim 1, is characterized in that: described working electrode is glass-carbon electrode.
3. a preparation method for the solid-state electrochemistry illumination sensor for detection of mercury ion according to claim 1, is characterized in that concrete steps are as follows:
(1) the immobilized preparation that has DNA1 glass-carbon electrode
A. by diameter, be the alundum (Al2O3) polishing that the glass-carbon electrode of 3～5 mm is used 1.0 μ m, 0.3 μ m, 0.05 μ m successively, then use successively ethanol, water ultrasonic cleaning, after water is rinsed well, nitrogen dries up standby;
B. step (a) gained glass-carbon electrode is placed in to the H containing N-(4-ammonia butyl)-different luminol of N-ethyl (ABEI) 2sO 4in solution, carry out cyclic voltammetry scan, voltage range is 0.2～1.5 V, with fast scan round 20～30 circles of sweeping of 0.01～0.1 V/s, makes ABEI in the polymerization of glass-carbon electrode surface electrical; The described H containing N-(4-ammonia butyl)-different luminol of N-ethyl (ABEI) 2sO 4n-in solution (4-ammonia the butyl)-different luminol of N-ethyl (ABEI) concentration is 0.01～0.0001 mol/L, H 2sO 4concentration is 0.1～1 mol/L;
C., after 0.01～0.1 M PBS damping fluid of pH=7～8 is rinsed well for glass-carbon electrode by step (b) gained, on glass-carbon electrode surface, drip 1～3 wt% glutaraldehyde solution 10～20 μ L, standing 20～40 minutes;
D. after by step (c) gained, for glass-carbon electrode, 0.01～0.1 M PBS damping fluid of pH=7～8 is rinsed well, on glass-carbon electrode surface, drip 0.01～0.1 M PBS damping fluid 10～20 μ L containing pH=7～8 of 10 μ mol/L DNA1, standing 20～40 min, 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 μ L mass percentage concentration are 2%, seals 1～2 h, obtain the glass-carbon electrode of the immobilized DNA1 of having;
(2) assembling of solid-state electrochemistry illumination sensor
A. prepare a series of variable concentrations containing the standard solution of mercury ion, by standard solution with contain 10 μ mol/L DNA2 pH=7～8 0.01～0.1 M PBS damping fluid by volume 1:1 mix;
B. get mixed solution 10～20 μ L of step (a) gained, be added drop-wise to the glass-carbon electrode surface of the immobilized DNA1 of having, under room temperature, hatch after 20～30 min, by 0.01～0.1 M PBS buffer solution for cleaning of pH=7～8;
C. the Avidin solution of getting 0.01～0.1 M PBS damping fluid preparation 0.01～0.1 mg/mL of appropriate pH=7～8, is added drop-wise to step (b) gained glass-carbon electrode surface by 10～20 μ L Avidin solution, hatches after 5 min;
D. by after 0.01～0.1 M PBS buffer solution for cleaning of pH=7～8 for glass-carbon electrode of step (c) gained, obtain the solid-state electrochemistry illumination sensor for mercury ion detecting.
4. the preparation method of a kind of solid-state electrochemistry illumination sensor for detection of mercury ion according to claim 3, is characterized in that: the structural formula of described DNA1 is: 5 '-NH 2-(CH 2) 6-GAC tgTC tcGT tcGC ttAG-3 '; The structural formula of described DNA2 is: 5 '-biotin-CTA tgCG taCG tgAC tgTC-3 '.
5. according to a detection method for the solid-state electrochemistry illumination sensor of the detection mercury ion described in any one in claim 1-4, it is characterized in that concrete steps are as follows:
(1) typical curve is set up
A. after the glass-carbon electrode of the immobilized DNA1 of having claimed in claim 3 being rinsed well with 0.01～0.1 M PBS damping fluid, in 0.05～0.2 M carbonate buffer solution of pH=9～10, start electrochemical reaction, test electrochemiluminescence intensity I 0;
B. the solid-state electrochemistry illumination sensor for mercury ion detecting claimed in claim 3 is put into 0.05～0.2 M carbonate buffer solution of pH=9～10, started electrochemical reaction, measure electrochemiluminescence intensity, obtain the Hg of a series of variable concentrations 2+the electrochemiluminescence intensity level I that solution is corresponding 1, calculate the Hg of a series of variable concentrations 2+the change value Δ I=I of solution electrochemistry luminous intensity 0i 1, set up change value Δ I and the Hg of electrochemiluminescence intensity 2+quantitative relationship between solution concentration;
(2) testing sample is measured
A. by containing the testing sample solution of mercury ion with contain 10 μ mol/L DNA2 pH7～8 0.01～0.1 M PBS damping fluid by volume 1:1 mix;
B. get mixed solution 10～20 μ L of step (a) gained, be added drop-wise to the glass-carbon electrode surface of the immobilized DNA1 of having, under room temperature, hatch after 20～30 min, by 0.01～0.1 M PBS buffer solution for cleaning of pH=7～8;
C. the Avidin solution of 10～20 μ, 0.01～0.1 mg/mL of 0.01～0.1 M PBS damping fluid preparation of pH=7～8 for L is added drop-wise to step (b) gained glass-carbon electrode surface, hatches after 5 min;
D. by after 0.01～0.1 M PBS buffer solution for cleaning of pH=7～8 for glass-carbon electrode of step (c) gained, as working electrode, saturated calomel electrode or Ag/AgCl electrode are contrast electrode, platinum electrode is to electrode, build three-electrode system, insert in 0.05～0.2 M carbonate buffer solution of pH=9～10, start electrochemical reaction, test electrochemiluminescence intensity level, according to change value Δ I and the Hg of electrochemiluminescence intensity 2+quantitative relationship between solution concentration, calculates Hg in testing sample solution 2+accurate concentration C hg.
6. a kind of detection method that detects the solid-state electrochemistry illumination sensor of mercury ion according to claim 5, is characterized in that: in described carbonate buffer solution, containing the hydrogen peroxide of 1 mM, the electrochemical method of employing is potential step chronoamperometry; Potential step: 0 V step to 1 V; Pulse width: 0.25 s; Measuring intervals of TIME: 30 s.
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