CN103913496B - A kind of preparation method of modified glassy carbon electrode and application thereof - Google Patents
A kind of preparation method of modified glassy carbon electrode and application thereof Download PDFInfo
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Abstract
The invention discloses a kind of Ru (bpy)3 2+The preparation method of-golden nanometer particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode, particularly, comprising: the polishing of glass-carbon electrode; Apply Ru (bpy)3 2+, preparation Ru (bpy)3 2+Modified glassy carbon electrode; Coated with gold nano particle, preparation Ru (bpy)3 2+-golden nanometer particle modified glassy carbon electrode; Apply single stranded DNA, preparation Ru (bpy)3 2+-golden nanometer particle-DNA modification glass-carbon electrode, wherein said single stranded DNA sequence is rich in base T, in conjunction with Hg2+After can complementary form stable double-stranded DNA structure, and 5 ' end of described single stranded DNA sequence is connected with SH-(CH2)6Group; Apply ferrocene functionalization graphene, preparation Ru (bpy)3 2+-golden nanometer particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode. Modified glassy carbon electrode prepared by the present invention can be used for detecting mercury ion content in water sample, has good electrochemical luminescence performance and excellent stability and reappearance, highly sensitive, simple to operation.
Description
Technical field
The present invention relates to electrochemical luminescence field, be specifically related to a kind of Ru (bpy)3 2+Preparation method and the application thereof of-golden nanometer particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode.
Background technology
Mercury is a kind of toxic metal element being distributed widely in environment, is one of conventional heavy metal of the industry such as battery, mining. Mercury can pass through the different approaches such as respiratory tract, skin or alimentary canal and invade human body, and food chain has extremely strong accumulation ability for mercury, and the toxicity of mercury accumulates, taking chronic as common, taking spirit-dysautonomia, gingivitis, tremble as cardinal symptom. Therefore, the qualitative detection of mercury ion and quantitative analysis all have great importance to life, environment and medical science and industrial and agricultural production etc.
At present, in the aqueous solution, the detection of mercury ion and the conventional method of quantitative analysis have: AAS, atomic absorption spectrum, plasma emission spectrum, Cold Atomic Fluorescent Mercury spectrum, gas-chromatography etc., detecting principle is that Photoinduced Electron shifts (PET), intramolecular charge shifts (ICT) and chemical reaction system etc., although the sensitivity of these methods is very high, but required instrument and equipment and analysis cost costliness, trivial operations, length consuming time, can not meet the demand of simple and quick detection Trace Hg. Emerging biology sensor method (enzyme inhibition method), in the time detecting mercury ion, have advantage quick, easy, with low cost, but detection limit is higher, and linear response range is narrow.
Electrochemical luminescence (Electrochemiluminescence, ECL) be the coefficient result of chemiluminescence and electrochemistry, refer to that ground state molecule is by participating in after electrochemical reaction obtains energy transitting to excitation state, luminous phenomenon when excitation state returns ground state, there is checkout gear simple, highly sensitive, can control the advantages such as the extent of reaction, but current electrochemical sensor is often lacking selectively aspect detection mercury ion. DNA biology sensor is using nucleic acid as molecular recognition and detection layers combining and a kind of trace analysis that can carry out fast detecting, monitoring in material molecule level of producing with the transducer of various physics, chemistry. DNA biology sensor is in the advantage of the aspects such as stability, repeatability, rapidity, convenience, sensitivity. It is a very fast research field of development in recent years that electrochemical method is applied to DNA biology sensor. Large biological molecule DNA and chemotron (being the electrode of current mode or electric potential type) combination can form DNA electrochemica biological sensor. Sensors with auxiliary electrode were has that electrode fabrication is easy, long service life, reproducible, highly sensitive, cost is low, less energy consumption, portable, not damage test sample, be not subject to solution colour to affect, be easy to realize the plurality of advantages such as microminiaturized.
Summary of the invention
The object of the present invention is to provide a kind of rapid sensitive to detect preparation method and the application thereof of the DNA electrochemica biological sensor (being electrode) of trace amount mercury ion in water sample.
On the one hand, the invention provides a kind of preparation method of modified glassy carbon electrode, it is characterized in that, comprise the following steps:
(1) glass-carbon electrode is carried out to pretreatment;
(2) tris (bipyridine) ruthenium (Ru (bpy)3 2+) apply: by Ru (bpy)3 2+Be coated to the described pretreated glass-carbon electrode surface of step (1), form Ru (bpy)3 2+Modified glassy carbon electrode;
(3) golden nanometer particle applies: golden nanometer particle is coated to the Ru (bpy) that step (2) obtains3 2+The glass-carbon electrode surface of modifying, forming surface coverage has the Ru (bpy) of Au nanoparticles films3 2+-golden nanometer particle modified glassy carbon electrode;
(4) DNA applies: single stranded DNA is coated to the Ru (bpy) that step (3) obtains3 2+-golden nanometer particle modified glassy carbon electrode surface, forms Ru (bpy)3 2+-golden nanometer particle-DNA modification glass-carbon electrode;
(5) ferrocene functionalization graphene applies: ferrocene functionalization graphene is coated to the Ru (bpy) that step (4) obtains3 2+-golden nanometer particle-DNA modification glass-carbon electrode surface, forms Ru (bpy)3 2+-golden nanometer particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode;
Wherein, described single stranded DNA sequence is rich in base T, in conjunction with Hg2+After can complementary form stable double-stranded DNA structure, and 5 ' end of described single stranded DNA sequence is connected with SH-(CH2)6Group.
In a specific embodiment of the present invention, the Ru (bpy) that step (2) is described3 2+Apply and carry out in the following manner: pretreated glass-carbon electrode step (1) Suo Shu is put into 0.1～1.0mMRu (bpy)3 2+With 0.01～0.1MKNO3Mixed solution in, to carry out potentiostatic scanning through pretreated glass-carbon electrode in the three-electrode system of working electrode, wherein, voltage is 1.8～2.0V, be 5～10min sweep time; Take out after electrode, wash by deionized water; Drying at room temperature.
In a specific embodiment of the present invention, the described golden nanometer particle of step (3) applies and carries out in the following manner: the Ru (bpy) that step (2) is obtained3 2+The glass-carbon electrode of modifying is put into 0.01～0.1M chlorauric acid solution, with Ru (bpy)3 2+Modified glassy carbon electrode is in the three-electrode system of working electrode, controls current potential at-1.0～1.1V, with chronoamperometry electro-deposition 5～20s; Take out after electrode, wash by deionized water; Drying at room temperature.
In a specific embodiment of the present invention, the described DNA of step (4) applies and carries out in the following manner: the Ru (bpy) that step (3) is obtained3 2+-golden nanometer particle modified glassy carbon electrode immerses in 5～10 μ MDNA solution and soaks 3～3.5h; Then, in 1～5mM mercaptoethanol solution, process 45～60min; Wash with phosphate buffer.
In a specific embodiment of the present invention, the described ferrocene functionalization graphene of step (5) applies and carries out in the following manner: the Ru (bpy) that step (4) is obtained3 2+-golden nanometer particle-DNA modification glass-carbon electrode immerses 1～2mgmL-145～60min in ferrocene functionalization graphene solution; Wash with phosphate buffer.
In a specific embodiment of the present invention, the described pretreatment of glassy carbon electrode of step (1) carries out in the following manner: by α-A1 for glass-carbon electrode2O3Polishing powder polishing; Use deionized water washes clean; Ultrasonic 10min in deionized water and ethanol respectively.
On the other hand, the present invention also provides and has utilized the made modified glassy carbon electrode of the method for this law in the application that detects mercury ion content in water sample.
In a specific embodiment of the present invention, modified glassy carbon electrode of the present invention comprises the following steps in the application that detects mercury ion in water sample:
1) by Ru (bpy)3 2+-golden nanometer particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode immerses 25～35min in water sample to be detected, then cleans described electrode with phosphate buffer;
2) by Ru (bpy)3 2+In the three-electrode system of-golden nanometer particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode as working electrode, using 0.05～0.1M Tri-n-Propylamine solution, as detecting, liquid carries out electrochemistry and electrochemical luminescence detects.
The present invention carries out Ru (bpy) on glass-carbon electrode surface3 2+Modify with Jenner's grain of rice, again by carrying out constructed dna electrochemical luminescence biology sensor at modified glassy carbon electrode surface self-organization single stranded DNA and ferrocene functionalization graphene, i.e. Ru (bpy)3 2+-golden nanometer particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode. This DNA electrochemical luminescence biology sensor utilizes single stranded DNA to realize ferrocene (Fc) to Ru (bpy) to the absorption of Graphene uniqueness3 2+The cancellation of electrochemistry optical signalling, due to Hg2+Can with single stranded DNA sequence in T-T base-pair generation specific binding form stable T-Hg2+-T complex structure, thus single stranded DNA occurred conformation is changed, form stable duplex structure, no longer adsorb Graphene, ferrocene Graphene is departed from, Ru in sensor (bpy) from DNA chain3 2+Electrochemistry optical signalling is recovered, and cancellation and the recovery of the electrochemistry optical signalling by sensor, realize the trace heavy metal contaminated ion Hg in visual testing environment water quality2+. Compared with prior art, the DNA electrochemical luminescence biology sensor tool of the present invention design has the following advantages: 1) have good stability, reappearance, mercury ion is had very high selective; 2) Graphene and golden nanometer particle are modified in electrode surface, the characteristic such as good conduction, high-specific surface area of having utilized Graphene and Jenner's grain of rice to have, thus increase greatly the sensitivity of electrode; 3) simple to operation, can quick and precisely detect the trace amount mercury ion in water sample. Therefore, DNA electrochemical luminescence biology sensor heavy metal ions in testing environment water quality of the present invention's design embodies good development prospect.
Brief description of the drawings
Fig. 1 be modified glassy carbon electrode of the present invention for detection of mercury ion experimental principle figure.
Fig. 2 is the cyclic voltammogram of different glass-carbon electrodes in an embodiment.
Fig. 3 be one embodiment of the invention modified glassy carbon electrode the different modifying stage electrochemical impedance.
Fig. 4 shows Hg2+The impact of concentration and the binding time electrochemical luminescence intensity on embodiment of the present invention modified glassy carbon electrode.
Fig. 5 shows the impact of pH value and the electrochemical luminescence intensity of salinity on embodiment of the present invention modified glassy carbon electrode.
Fig. 6 is that the modified glassy carbon electrode of one embodiment of the invention is at the electrochemical luminescence signals in different modifying stage.
Fig. 7 is that the modified glassy carbon electrode of one embodiment of the invention detects variable concentrations Hg2+Electrochemical luminescence signals time, electrochemical luminescence intensity and Hg2+The canonical plotting of concentration.
Fig. 8 is the modified glassy carbon electrode anti-interference resolution chart of one embodiment of the invention.
Detailed description of the invention
Electrochemical luminescence detects, and is a kind of new analytical method growing up on chemiluminescent basis, is the product that chemiluminescence and electrochemistry mutually combine, and has high sensitivity, high selectivity, is easy to realize the features such as automatic continuous analysis. Ru (bpy)3 2+Owing to thering is the good characteristic that is stimulated, be day by day subject to people's attention in electrochemical luminescence field. Ferrocene and derivative thereof are the important object that coordination structure chemistry and inorganic photochemical are studied always, the structure that it is special and to Ru (bpy)3 2+Act in basic research significant Deng the cancellation of sensitising agent.
Nanogold particle has unique physics, chemical property and good bio-compatibility, has been widely used in DNA electrochemica biological sensor. The application of golden nanometer particle in electrochemica biological sensor is mainly aspect two: the one, improve can molecular recognition material process for fixation, improve fixed amount; The 2nd, the sensitivity that improves electrochemical signals indicator, thereby the detection sensitivity of DNA electrochemica biological sensor, shorten detection time and improve and detect flux.
Graphene is the crystal of banishing honeycomb lattice structure of the tightly packed formation of monolayer carbon atom, and unique structure makes it have excellent electricity, calorifics, mechanics and chemical property, makes it can be used as novel sensor material. Graphene can adsorb single stranded DNA effectively by pi-pi accumulation effect, but because pi-pi accumulation effect can not occur effectively for the rigid structure of double-stranded DNA itself and the hydrophily of outer both sides phosphoric acid skeleton, so Graphene can not adsorb double-stranded DNA effectively.
The ingenious interaction relationship having utilized between above-mentioned substance of the present invention, designs a kind of Ru (bpy) that detects mercury ion that can be used in3 2+-golden nanometer particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode, the principle of its detection mercury ion as shown in Figure 1. There is no Hg2+Time, single stranded DNA absorption Graphene, thus realize ferrocene to Ru (bpy)3 2+The cancellation of electrochemistry optical signalling; There is Hg2+Time, due to Hg2+Can with single stranded DNA sequence in T-T base-pair generation specific binding form stable T-Hg2+-T complex structure, thus single stranded DNA occurred conformation is changed, form stable duplex structure, no longer adsorb Graphene, ferrocene Graphene is departed from, Ru in sensor (bpy) from double-stranded DNA3 2+Electrochemistry optical signalling is recovered. By cancellation and the recovery of electrochemistry optical signalling, and show intensity, can realize trace Hg in water sample2+Detection.
The parameter such as electrode reaction character, mechanism and kinetics of electrode process that detects mercury ion in order to study modified glassy carbon electrode of the present invention, pretreated glass-carbon electrode, before modifying, adopts three-electrode system cyclic voltammetry to detect. In one embodiment, be pretreated glass-carbon electrode by working electrode, be platinum electrode to electrode, reference electrode is the three-electrode system of Ag/AgCl electrode, in 0.5M sulfuric acid, glass-carbon electrode is carried out to cyclic voltammetry scan, it is-0.2～1.6V that voltage is wherein set, and after detection, uses deionized water rinsing glass-carbon electrode, dry up glass-carbon electrode surface, for subsequent use. Similarly, at DNA electrochemica biological sensor, i.e. Ru (bpy)3 2+Time prepared by-golden nanometer particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode, also need to carry out cyclic voltammetry scan, in one embodiment, the modified glassy carbon electrode that preparation is completed is put into 0.05～0.1M Tri-n-Propylamine solution and is carried out cyclic voltammetry scan, wherein voltage range is 0.2～1.3V, and sweep speed is 50～100mVs-1, photomultiplier high pressure is set to-800～-1000V.
Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are described, should be appreciated that preferred embodiment described herein, only for description and interpretation the present invention, is not intended to limit the present invention.
Embodiment 1
The present invention uses electrochemical workstation and MPI-B type multi-parameter chemiluminescence analysis test macro.
1. pretreatment of glassy carbon electrode: glass-carbon electrode is through α-A1 of 0.05 μ m2O3After polishing powder polishing, rinse well by deionized water, and ultrasonic 10min in deionized water and ethanol respectively. Employing three-electrode system detects, working electrode is glass-carbon electrode, be platinum electrode to electrode, reference electrode is Ag/AgCl electrode, in 0.5M sulfuric acid, it is-0.2～1.6V that voltage is set, glass-carbon electrode is carried out to cyclic voltammetry scan, after detection, use deionized water rinsing electrode, dry up electrode surface, for subsequent use.
2. the preparation of modified glassy carbon electrode:
1) pretreatment rear electrode is put into 0.1mMRu (bpy)3 2+And 0.01MKNO3Mixed solution in, on electrochemical workstation, control anode potential and process 5min at 1.8V place constant potential, take out electrode, with deionized water washing, drying at room temperature; Electrode is put into 0.01M chlorauric acid solution, on electrochemical workstation, control current potential at-1.0～1.1V chronoamperometry electro-deposition 5s, take out electrode, by deionized water washing, drying at room temperature.
2) modified electrode step 1) being obtained immerses in 10 μ M single stranded DNA solution and soaks 3h, then in 1mM mercaptoethanol solution, processes 60min, cleans modified glassy carbon electrode with phosphate buffer; Just glass-carbon electrode immerses 2mgmL again-160min in ferrocene functionalization graphene solution, uses phosphate buffer cleaning electrode, obtains Ru (bpy)3 2+-golden nanometer particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode.
Wherein, the nucleotides sequence of single stranded DNA is classified 5 ' ATTCTTTGTTCTCCCCTGTTCTTTGTTT-3 ' as, is synthesized by biotech firm. In order to strengthen the connection between DNA and golden nanometer particle, in the time of synthesizing single-stranded DNA, be connected with mercapto groups (SH-(CH at its 5 ' end2)6), connect DNA and golden nanometer particle by sulfydryl-Au effect.
Embodiment 2
1. pretreatment of glassy carbon electrode: glass-carbon electrode is through α-A1 of 0.3 μ m2O3After polishing powder polishing, rinse well by deionized water, and ultrasonic 10min in deionized water and ethanol respectively. Employing three-electrode system detects, working electrode is glass-carbon electrode, be platinum electrode to electrode, reference electrode is Ag/AgCl electrode, in 0.5M sulfuric acid, it is-0.2～1.6V that voltage is set, glass-carbon electrode is carried out to cyclic voltammetry scan, after detection, use deionized water rinsing electrode, dry up electrode surface, for subsequent use.
2. the preparation of modified glassy carbon electrode:
1) pretreatment rear electrode is put into 1.0mMRu (bpy)3 2+And 0.1MKNO3Mixed solution in, on electrochemical workstation, control anode potential and process 10min at 2.0V place constant potential, take out electrode, with deionized water washing, drying at room temperature; Electrode is put into 0.1M chlorauric acid solution, on electrochemical workstation, control current potential at-1.0～1.1V chronoamperometry electro-deposition 20s, take out electrode, by deionized water washing, drying at room temperature.
2) modified electrode step 1) being obtained immerses in 5 μ M single stranded DNA solution and soaks 3.5h, then in 5mM mercaptoethanol solution, processes 60min, cleans modified glassy carbon electrode with phosphate buffer; Just glass-carbon electrode immerses 1mgmL again-145min in ferrocene functionalization graphene solution, uses phosphate buffer cleaning electrode, obtains Ru (bpy)3 2+-golden nanometer particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode.
Wherein, the nucleotides sequence of single stranded DNA is classified 5 ' ATTCTTTGTTCTCCCCTGTTCTTTGTTT-3 ' as, is synthesized by biotech firm. In order to strengthen the connection between DNA and golden nanometer particle, in the time of synthesizing single-stranded DNA, be connected with mercapto groups (SH-(CH at its 5 ' end2)6), connect DNA and golden nanometer particle by sulfydryl-Au effect.
The Performance Detection of modified glassy carbon electrode
1) cyclic voltammetry detects
Fig. 2 is the cyclic voltammogram of different glass-carbon electrodes, and wherein a is glass-carbon electrode bare electrode, and b is gold nano-glass-carbon electrode, and c is Ru (bpy)3 2+-glass-carbon electrode, d is Ru (bpy)3 2+-gold nano-glass-carbon electrode. At 0.01MKNO3In solution, carry out cyclic voltammetry scan, curve a is very level and smooth, does not have obvious redox peak to occur; After electrode coated with gold nano particle, there is oxidation peak at 0.922V in curve b, illustrates that golden nanometer particle is modified at glass-carbon electrode surface effectively; Electrode is applied to Ru (bpy)3 2+After, having there is an obvious oxidation peak at 1.01V in curve c, shows Ru (bpy)3 2+Effectively be modified at glass-carbon electrode surface; Again at Ru (bpy)3 2+Modified electrode coated with gold nano particle, preparation Ru (bpy)3 2+-golden nanometer particle modified glassy carbon electrode, there is an oxidation peak that signal strengthens at 1.05V in curve d, this shows that golden nanometer particle can effectively strengthen electronics transmission efficiency, thereby makes Ru (bpy)3 2+Signal peak be enhanced.
2) electrochemical impedance detects
Fig. 3 is the electrochemical impedance of the glass-carbon electrode in different modifying stage. Wherein, curve a is the nyquist plot of blank glass-carbon electrode, as seen from the figure, and at 5.0mM[Fe (CN)6]3?/4?In system, blank glass-carbon electrode has presented very little radian at high frequency region, and the nyquist plot obtaining is almost straight line, and this has showed typical blank electrode surface swift electron transmittance process. Electrode is applied to Ru (bpy)3 2+After golden nanometer particle, the curve radian of curve b high frequency region obviously increases, and can be interpreted as Ru (bpy)3 2+Thereby cause a non-equal face boundary of electrode to produce the electron transfer characteristic that has changed electrode surface with golden nanometer particle to the modification of electrode. After single stranded DNA and ferrocene functionalization graphene modified electrode, the conductance property of electrode strengthens to some extent, and as shown in curve c, its high frequency region camber ratio curve b reduces to some extent, illustrates that ferrocene functionalization graphene can effectively promote electronics transmission. As Ru (bpy)3 2+-golden nanometer particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode combines Hg2+Afterwards, curve d high frequency region curve radian increases to some extent, illustrates that ferrocene functionalization graphene has departed from DNA surface.
3）Hg2+The impact on electrochemical luminescence intensity of concentration and binding time
Fig. 4 is the Ru (bpy) of preparation3 2+-golden nanometer particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode is respectively at the Hg of 0.05nM, 1nM and 100nM2+The electroluminescence intensity level obtaining during in conjunction with different time in solution, thus draw electrochemical luminescence intensity and binding time, Hg2+Interaction relationship chart between concentration three. As can be seen from Figure 4, electrochemical luminescence intensity and binding time, Hg2+There is good linear relationship in concentration.
4) impact of water environment on electrochemical luminescence intensity
Fig. 5 has shown that pH value (A) and sodium chloride concentration (B) are to Ru (bpy)3 2+The impact of-golden nanometer particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode. As seen from Figure 5, be that 7.5 left and right and sodium chloride concentration can be realized electrode sensor to Hg while being 0.1M at pH2+Optimum detection.
5) electrode sensor stability study
Fig. 6 shown in 0.1M Tri-n-Propylamine solution Different electrodes carried out to electrochemical luminescence intensity detection, and within a certain period of time, the electrochemical luminescence intensity of sensor does not have considerable influence, more stable, and wherein a is glass-carbon electrode bare electrode; B is Ru (bpy)3 2+-glass-carbon electrode; C is Ru (bpy)3 2+-gold nano-single stranded DNA-ferrocene functionalization graphene modified glassy carbon electrode; D is Ru (bpy)3 2+-gold nano-double-stranded DNA-ferrocene functionalization graphene modified glassy carbon electrode. As seen from Figure 6, in the time that voltage is 1.1～1.2V, there is Ru (bpy)3 2+Electrochemistry light intensity signal occur. The time of Fig. 6 mid portion and electrochemical luminescence strength relationship chart show, in 0.1M Tri-n-Propylamine solution, modified electrode is carried out to continuous sweep, its electrochemical luminescence intensity does not have considerable influence, more stable, illustrate that electrode sensor of the present invention has good stability.
6) mercury ion detecting
Utilize the modified glassy carbon electrode of preparation to detect mercury ion in water quality, concrete detecting step is as follows:
1) modified glassy carbon electrode is immersed to the Hg of variable concentrations to be detected2+30min in solution, then uses phosphate buffer cleaning electrode.
2), using modified glassy carbon electrode as working electrode, on electrochemical workstation, use 0.1M Tri-n-Propylamine solution to carry out electrochemical luminescence detection as detecting liquid. (cyclic voltammetry scan voltage range is 0.2～1.25V, and sweep speed is 100mVs-1, be set to-800V of photomultiplier high pressure).
Fig. 7 has shown the Hg of modified glassy carbon electrode at variable concentrations2+(Hg in solution2+Concentration sequence is 0.05nm(a), 0.1nm(b), 1nm(c) and, 10nm(d), 20nm(e) and, 50nm(f) and 100nm(g)) cultivate after certain hour the electrochemical luminescence intensity vs time graph in 0.1M Tri-n-Propylamine solution. As seen from the figure, under best experiment condition, work as Hg2+Concentration is between 0.05～100nM time, along with Hg2+The variation of concentration, electrochemical luminescence intensity has significant change. Detect Hg through calculating2+Linear equation be ΔI ECL=371lgC Hg2+ + 4226.6, coefficient correlation is 0.9968, detects and is limited to 18pM.
7) anti-interference test
For probing into the interference free performance of sensor to other potential interference ions, we have carried out parallel disturbed test experiment to the different ions that may coexist with mercury ion. In the detection shown in Fig. 8, ion concentration of mercury is 10nM, and other ion concentrations are 500nM. As seen from Figure 8, the mercury ion of 50 times of dilutions has produced ten response signals times over interfering ion, illustrates that the sensor of this experimental design has good Hg2+Identification selection.
Finally it should be noted that: the foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, although the present invention is had been described in detail with reference to previous embodiment, for a person skilled in the art, its technical scheme that still can record aforementioned each embodiment is modified, or part technical characterictic is wherein equal to replacement. Within the spirit and principles in the present invention all, any amendment of doing, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (8)
1. a preparation method for modified glassy carbon electrode, is characterized in that, comprises the following steps:
(1) glass-carbon electrode is carried out to pretreatment;
(2)Ru(bpy)3 2+Apply: by Ru (bpy)3 2+Be coated to described pretreated of step (1)Glass-carbon electrode surface, forms Ru (bpy)3 2+Modified glassy carbon electrode;
(3) golden nanometer particle applies: golden nanometer particle is coated to the Ru (bpy) that step (2) obtains3 2+RepairThe glass-carbon electrode surface of decorations, forming surface coverage has the Ru (bpy) of Au nanoparticles films3 2+-golden nanometer particleModified glassy carbon electrode;
(4) DNA applies: single stranded DNA is coated to the Ru (bpy) that step (3) obtains3 2+-golden nanometer particle is repaiiedDecorations glass-carbon electrode surface, forms Ru (bpy)3 2+-golden nanometer particle-DNA modification glass-carbon electrode;
(5) ferrocene functionalization graphene applies: ferrocene functionalization graphene is coated to step (4)The Ru (bpy) obtaining3 2+-golden nanometer particle-DNA modification glass-carbon electrode surface, forms Ru (bpy)3 2+-JinNano particle-DNA-ferrocene functionalization graphene modified glassy carbon electrode;
Wherein, described single stranded DNA sequence is rich in base T, in conjunction with Hg2+After can complementary form stable two strandsDNA structure, and 5 ' end of described single stranded DNA sequence is connected with SH-(CH2)6Group.
2. the preparation method of modified glassy carbon electrode as claimed in claim 1, wherein described in step (2)Ru (bpy)3 2+Apply and carry out in the following manner: by pretreated glass carbon step (1) Suo ShuElectrode is put into 0.1～1.0mMRu (bpy)3 2+With 0.01～0.1MKNO3Mixed solution in, withIn the three-electrode system that is working electrode through pretreated glass-carbon electrode, carry out potentiostatic scanning, itsIn, voltage is 1.8～2.0V, be 5～10min sweep time; Washing; Dry.
3. the preparation method of modified glassy carbon electrode as claimed in claim 1, wherein described in step (3)Golden nanometer particle apply carry out in the following manner: the Ru (bpy) that step (2) is obtained3 2+ModifyGlass-carbon electrode is put into 0.01～0.1M chlorauric acid solution, with Ru (bpy)3 2+Modified glassy carbon electrode isIn the three-electrode system of working electrode, control current potential at-1.0～1.1V, use chronoamperometry electro-deposition5～20s; Washing; Dry.
4. the preparation method of modified glassy carbon electrode as claimed in claim 1, wherein described in step (4)DNA apply carry out in the following manner: the Ru (bpy) that step (3) is obtained3 2+-golden nanometer particle is repaiiedDecorations glass-carbon electrode immerses in 5～10 μ MDNA solution and soaks 3～3.5h; Then, at 1～5mM sulfydrylIn ethanolic solution, process 45～60min; Wash with phosphate buffer.
5. the preparation method of modified glassy carbon electrode as claimed in claim 1, wherein described in step (5)Ferrocene functionalization graphene apply carry out in the following manner: step (4) is obtainedRu(bpy)3 2+-golden nanometer particle-DNA modification glass-carbon electrode immerses 1～2mgmL-1Ferrocene function fossil45～60min in China ink alkene solution; Wash with phosphate buffer.
6. the preparation method of modified glassy carbon electrode as claimed in claim 1, wherein described in step (1)Pretreatment of glassy carbon electrode carry out in the following manner: by α-A1 for glass-carbon electrode2O3Polishing powder polishing;Washing; Ultrasonic 10min in deionized water and ethanol respectively.
7. the modified glassy carbon electrode that as described in as arbitrary in claim 1～6, method is made is detecting in water sampleThe application of mercury ion content.
8. application as claimed in claim 7, is characterised in that, comprises the following steps:
1) by described Ru (bpy)3 2+-golden nanometer particle-DNA-ferrocene functionalization graphene is modified glass carbon electricityThe utmost point immerses 25～35min in water sample to be detected, then cleans described electrode with phosphate buffer;
2) by described Ru (bpy)3 2+-golden nanometer particle-DNA-ferrocene functionalization graphene is modified glass carbon electricityIn the three-electrode system of the utmost point as working electrode, using 0.05～0.1M Tri-n-Propylamine solution as detectionLiquid carries out electrochemistry and electrochemical luminescence detects.
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