CN103558275A - Method for detecting mercury ions by nucleic acid aptamer based photoelectrochemistry - Google Patents
Method for detecting mercury ions by nucleic acid aptamer based photoelectrochemistry Download PDFInfo
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Abstract
The invention relates to a method for detecting mercury ions by nucleic acid aptamer based photoelectrochemistry. An On-Off type nucleic acid aptamer based photoelectrochemical sensor is adopted for detecting the mercury ions with different concentrations and is constructed based on quercetin copper sensitization perylene tetracarboxylic-oxidized graphene heterojunction. Based on double advantages of a quercetin copper compound as a photosensitizer and a reducing agent, the method can be used for double amplifying photoelectrical signals, and has the characteristics of being convenient, simple, super sensitive, economical, high in specificity and the like. The constructed On-Off type nucleic acid aptamer based photoelectrochemical sensor adopts current-time technology to detect the mercury ions with different concentrations in a 0.1 mol/L phosphate buffered solution at the offset electric potential of 0.2V and under optical radiation of more than 450 nm.
Description
Technical field
The present invention relates to analytical test field, especially relate to a kind of method that can be applicable to the detection mercury ion of environmental monitoring.
Background technology
Mercury ion is one of common pollutant, and it can become hypertoxic methyl mercury by the role transformation of microorganism, by food chain, enter people's human body, causes mercury poisoning event.The method that detects at present mercury ion has, such as atomic spectroscopy, Molecular Spectroscopy, the chromatography of ions, electrochemical process and immunoassay etc.Yet every kind of method all has one of following shortcoming at least, as apparatus expensive, the feature such as method is complicated, consuming time, sensitivity is not high, reagent toxicity is large.Therefore invent new analytical approach and overcome especially necessity of above-mentioned shortcoming.It is a kind of method of testing of novelty that Optical Electro-Chemistry detects, the method is based on phot-luminescence telecommunications mark, to cause the separation of electron-hole pair, under suitable inclined to one side potential condition, realize electronics transmits fast on analyte, semiconductor and electrode, thereby formation photocurrent, through the analyte of recognition component specific binding, can affect quantitatively the variation of photocurrent, thereby realize, the Optical Electro-Chemistry of analyte be detected.With regard to amplifying signal, structure, dye-sensitized semiconductor, the plasma that the method for taking at present mainly contains metal or nonmetal doping semiconductor, heterojunction semiconductor strengthens the design of photosignal.Yet these methods are all consideration and design based on photoelectricity beacon aspect, suppress right compound of charge carrier, thereby strengthen photosignal and improve further Optical Electro-Chemistry detection sensitivity.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of method that aptamer based photoelectrochemicalcell detects mercury ion, its specific recognition mechanism based on aptamer, adopt Quercetin-Cu complex as the two-fold advantage of photosensitizer and reductive agent, can play the effect of dual amplification photosignal, and possess conveniently, the feature such as easy, hypersensitive, economy, high specific.
Aptamer based photoelectrochemicalcell detects a method for mercury ion, adopts " On-Off " type aptamer based photoelectrochemicalcell sensor of structure for detection of the mercury ion of variable concentrations; Described " On-Off " type aptamer based photoelectrochemicalcell sensor is the Optical Electro-Chemistry sensor based on Quercetin-Cu Minization perylene tetracarboxylic acid-graphene oxide heterojunction.It is the two-fold advantage as photosensitizer and reductive agent based on Quercetin-Cu complex, can play the effect of dual amplification photosignal, and possess conveniently, the feature such as easy, hypersensitive, economy, high specific." On-Off " type aptamer based photoelectrochemicalcell sensor building is in 0.1mol L
-1in phosphate buffered solution, at current potential 0.2V partially and be greater than the mercury ion that adopts electric current-time technology for detection variable concentrations under the optical radiation of 450nm.Can realize hypersensitive and the specific detection of mercury ion, detect and be limited to 3.3fmol L
-1.
Method of the present invention, the construction method of wherein said " On-Off " type aptamer based photoelectrochemicalcell sensor is:
(1) Bei perylene tetracarboxylic acid/graphene oxide processed heterojunction;
(2) Qu perylene tetracarboxylic acid/graphene oxide heterojunction solution drip and are applied to glass-carbon electrode surface and under room temperature, dry get perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode processed;
(3) amination T base (aptamers) is fixed on to perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode surface Hou EDC-NHS activation;
(4) then with its surface of TE damping fluid (Tris-HCl+1mmol/L EDTA) drip washing, then, by 0.25～1 μ mol L
-1amination T base soln drip and be coated with its electrode surface, after being incubated overnight at 4 ℃ with its surface of TE damping fluid drip washing; Then, get 0.25～1 μ mol L
-1its electrode surface of the molten drop-coated of Poly A, at 37 ℃ after incubation with its surface of TE damping fluid drip washing; Finally, by 5～20 μ mol L
-1quercetin-Cu ethanolic solution drip and be coated with its electrode surface, at 37 ℃ after incubation with its surface of TE damping fluid drip washing.
Method of the present invention, (a kind of enforceable preferred preparation method of 1) Zhong perylene tetracarboxylic acid/graphene oxide heterojunction is as follows: prepare 1 * 10 for wherein said step
-4mol L
-1perylene tetracarboxylic acid DMF solution and 1mg mL
-1graphene oxide solution, Ran Hou perylene tetracarboxylic acid solution and graphene oxide solution with the volume ratio of 5:1 fully mix, standing, i.e. Sheng Cheng perylene tetracarboxylic acid/graphene oxide heterojunction.
Method of the present invention, in wherein said step (3), a kind of enforceable preferred activation method of modified electrode is as follows: by 20 μ L5mmol L
-1eDC and 10mmol L
-1the Tris-HCl solution Di of the pH7.4 of NHS is surperficial Zhi perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode, and activates 1～2h under room temperature.
Method of the present invention has solved following technical matters:
The absorption of 1, expanding visible ray, has improved electricity conversion;
2, suppress right compound in semiconductor electronic-hole;
3, promote effective separation of electron-hole pair, improve the generation ratio of light induced electron;
4, build Quercetin-Cu Minization perylene tetracarboxylic acid-graphene oxide heterojunction based photoelectrochemicalcell sensing platform and use it for mercury ion hypersensitive and specific detection.
The Optical Electro-Chemistry sensing of method of the present invention based on Quercetin-Cu Minization perylene tetracarboxylic acid-graphene oxide heterojunction can strengthen significantly the absorption of visible ray and promote effective separation that charge carrier is right.Simultaneously this method possess that other advantage is as good in: hypersensitive, specificity, economy, environmental friendliness and low (the 3.3fmol L of detection limit
-1) etc. feature.
Accompanying drawing explanation
Fig. 1 is the making schematic diagram of Quercetin-Cu Minization perylene tetracarboxylic acid-graphene oxide heterojunction base light anode;
Fig. 2 is the photocurrent response of Quercetin-Cu Minization perylene tetracarboxylic acid-graphene oxide heterojunction base light anode to mercury ion; Hg
2+concentration range (from the top down): 0,0.01,0.02,0.06,0.08,0.1,0.4,0.6,0.8,1 and 2pmol L
-1; 0.1mol L
-1pH7.0 buffer salt solution; Bias potential 0.2V; Be greater than the 250W halogen tungsten lamp radiation of 450nm; Illustration is its working curve.
Embodiment
For further illustrating the present invention, illustrate with the following Examples:
Embodiment 1:
The preparation method of ， perylene tetracarboxylic acid/graphene oxide heterojunction is as follows as shown in Figure 1: first, prepare 2 * 10
-4mol L
-1perylene tetracarboxylic acid DMF solution and 2mg mL
-1graphene oxide solution, Ran Hou perylene tetracarboxylic acid solution and graphene oxide solution with the volume ratio of 10:1 fully mix, standing, and Sheng Cheng perylene tetracarboxylic acid/graphene oxide heterojunction.
Glass-carbon electrode, after 0.05 μ m particle diameter alundum (Al2O3) suspending liquid polishing, cleans up with acetone and pure water successively, then places room temperature and dries.Getting 10 μ L perylene tetracarboxylic acid/graphene oxide heterojunction solution drips and is applied to glass-carbon electrode surface and under room temperature, dries get perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode processed.Then, amination T base is fixed on to perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode surface Hou EDC-NHS activation.The concrete activation method of modified electrode is as follows: by 20 μ L10mmol L
-1eDC and 20mmol L
-1the Tris-HCl solution Di of the pH7.4 of NHS is surperficial Zhi perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode, and under room temperature, activates 2 hours.Then use its surface of TE damping fluid drip washing, then, by 10 μ L1 μ mol L
-1amination T base soln drip and be coated with its electrode surface, after being incubated overnight at 4 ℃ with its surface of TE damping fluid drip washing.Then, get 10 μ L1 μ mol L
-1its electrode surface of the molten drop-coated of Poly A, at 37 ℃ after incubation 1h with its surface of TE damping fluid drip washing.Finally, by 10 μ L20 μ mol L
-1quercetin-Cu ethanolic solution drip and be coated with its electrode surface, at 37 ℃ after incubation 1h with its surface of TE damping fluid drip washing.Utilize " T-Hg-T " coordination to form, the mercury ion of variable concentrations or interfering ion are dripped and be applied to electrode surface and at 37 ℃, after incubation 1h, use its surface of damping fluid drip washing again.The existence of mercury ion causes A-T chain to untie and form simultaneously " T-Hg-T ", untiing of A-T chain causes Quercetin-Cu to leave electrode surface, finally cause the decline of photocurrent, use just the existence of mercury ion to reduce quantitatively the mechanism of photocurrent, realize the structure of the adaptive based photoelectrochemicalcell sensor of " On-Off " type nucleic acid.The Optical Electro-Chemistry sensor of making is in 0.1mol L
-1in phosphate buffered solution, at current potential 0.2V partially and be greater than the mercury ion that adopts electric current-time technology for detection variable concentrations under the optical radiation of 450nm.
Embodiment 2:
The preparation method of perylene tetracarboxylic acid/graphene oxide heterojunction is as follows: first, prepare 1 * 10
-4mol L
-1perylene tetracarboxylic acid DMF solution and 1mg mL
-1graphene oxide solution, Ran Hou perylene tetracarboxylic acid solution and graphene oxide solution with the volume ratio of 5:1 fully mix, standing, and Sheng Cheng perylene tetracarboxylic acid/graphene oxide heterojunction.
Glass-carbon electrode, after 0.05 μ m particle diameter alundum (Al2O3) suspending liquid polishing, cleans up with acetone and pure water successively, then places room temperature and dries.Getting 5 μ L perylene tetracarboxylic acid/graphene oxide heterojunction solution drips and is applied to glass-carbon electrode surface and under room temperature, dries get perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode processed.Then, amination T base is fixed on to perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode surface Hou EDC-NHS activation.The concrete activation method of modified electrode is as follows: by 20 μ L5mmol L
-1eDC and 10mmol L
-1the Tris-HCl solution Di of the pH7.4 of NHS is surperficial Zhi perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode, and activates 1h under room temperature.Then use its surface of TE damping fluid drip washing, then, by 10 μ L0.5 μ mol L
-1amination T base soln drip and be coated with its electrode surface, after being incubated overnight at 4 ℃ with its surface of TE damping fluid drip washing.Then, get 10 μ L0.5 μ mol L
-1its electrode surface of the molten drop-coated of Poly A, at 37 ℃ after incubation 0.5h with its surface of TE damping fluid drip washing.Finally, by 10 μ L10 μ mol L
-1quercetin-Cu ethanolic solution drip and be coated with its electrode surface, at 37 ℃ after incubation 0.5h with its surface of TE damping fluid drip washing.Utilize " T-Hg-T " coordination to form, the mercury ion of variable concentrations or interfering ion are dripped and be applied to electrode surface incubation at 37 ℃ and after 0.5 hour, use its surface of damping fluid drip washing again.The existence of mercury ion causes A-T chain to untie and form simultaneously " T-Hg-T ", untiing of A-T chain causes Quercetin-Cu to leave electrode surface, finally cause the decline of photocurrent, use just the existence of mercury ion to reduce quantitatively the mechanism of photocurrent, realize the structure of the adaptive based photoelectrochemicalcell sensor of " On-Off " type nucleic acid.The Optical Electro-Chemistry sensor of making is in 0.1mol L
-1in phosphate buffered solution, at current potential 0.2V partially and be greater than the mercury ion that adopts electric current-time technology for detection variable concentrations under the optical radiation of 450nm.
Embodiment 3:
The preparation method of perylene tetracarboxylic acid/graphene oxide heterojunction is as follows: first, prepare 0.5 * 10
-4mol L
-1perylene tetracarboxylic acid DMF solution and 0.5mg mL
-1graphene oxide solution, Ran Hou perylene tetracarboxylic acid solution and graphene oxide solution with the volume ratio of 2.5:1 fully mix, standing, and Sheng Cheng perylene tetracarboxylic acid/graphene oxide heterojunction.
Glass-carbon electrode, after 0.05 μ m particle diameter alundum (Al2O3) suspending liquid polishing, cleans up with acetone and pure water successively, then places room temperature and dries.Getting 2.5 μ L perylene tetracarboxylic acid/graphene oxide heterojunction solution drips and is applied to glass-carbon electrode surface and under room temperature, dries get perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode processed.Then, amination T base is fixed on to perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode surface Hou EDC-NHS activation.The concrete activation method of modified electrode is as follows: by 10 μ L2.5mmol L
-1eDC and 5mmol L
-1the Tris-HCl solution Di of the pH7.4 of NHS is surperficial Zhi perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode, and activates 0.5h under room temperature.Then use its surface of TE damping fluid drip washing, then, by 5 μ L0.25 μ mol L
-1amination T base soln drip and be coated with its electrode surface, after being incubated overnight at 4 ℃ with its surface of TE damping fluid drip washing.Then, get 5 μ L0.25 μ mol L
-1its electrode surface of the molten drop-coated of Poly A, at 37 ℃ after incubation 0.25h with its surface of TE damping fluid drip washing.Finally, by 5 μ L5 μ mol L
-1quercetin-Cu ethanolic solution drip and be coated with its electrode surface, at 37 ℃ after incubation 0.25h with its surface of TE damping fluid drip washing.Utilize " T-Hg-T " coordination to form, the mercury ion of variable concentrations or interfering ion are dripped and be applied to electrode surface and at 37 ℃, after incubation 0.25h, use its surface of damping fluid drip washing again.The existence of mercury ion causes A-T chain to untie and form simultaneously " T-Hg-T ", untiing of A-T chain causes Quercetin-Cu to leave electrode surface, finally cause the decline of photocurrent, use just the existence of mercury ion to reduce quantitatively the mechanism of photocurrent, realize the structure of the adaptive based photoelectrochemicalcell sensor of " On-Off " type nucleic acid.The Optical Electro-Chemistry sensor of making is in 0.05mol L
-1in phosphate buffered solution, at current potential 0.2V partially and be greater than the mercury ion that adopts electric current-time technology for detection variable concentrations under the optical radiation of 450nm.
Fig. 2 is Quercetin-Cu Minization perylene tetracarboxylic acid-graphene oxide heterojunction base light anode of embodiment 1 photocurrent response to mercury ion; Hg
2+concentration range (from the top down): 0,0.01,0.02,0.06,0.08,0.1,0.4,0.6,0.8,1 and 2pmol L
-1; 0.1mol L
-1pH7.0 buffer salt solution; Bias potential 0.2V; Be greater than the 250W halogen tungsten lamp radiation of 450nm; Illustration is its working curve.
Above-described embodiment is described the preferred embodiment of the present invention; not scope of the present invention is limited; design under the prerequisite of spirit not departing from the present invention; various distortion and improvement that the common engineering technical personnel in this area make technical scheme of the present invention, all should fall in the definite protection domain of claims of the present invention.
Claims (6)
1. aptamer based photoelectrochemicalcell detects a method for mercury ion, it is characterized in that: adopt " On-Off " type aptamer based photoelectrochemicalcell sensor of structure for detection of the mercury ion of variable concentrations; Described " On-Off " type aptamer based photoelectrochemicalcell sensor is the Optical Electro-Chemistry sensor based on Quercetin-Cu Minization perylene tetracarboxylic acid-graphene oxide heterojunction.
2. method according to claim 1, is characterized in that: the construction method of described " On-Off " type aptamer based photoelectrochemicalcell sensor is:
(1) Bei perylene tetracarboxylic acid/graphene oxide processed heterojunction;
(2) Qu perylene tetracarboxylic acid/graphene oxide heterojunction solution drip and are applied to glass-carbon electrode surface and under room temperature, dry get perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode processed;
(3) amination T base (aptamers) is fixed on to perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode surface Hou EDC-NHS activation;
(4) then with its surface of TE damping fluid drip washing, then, by 0.25～1 μ mol L
-1amination T base soln drip and be coated with its electrode surface, after being incubated overnight at 4 ℃ with its surface of TE damping fluid drip washing; Then, get 0.25～1 μ mol L
-1its electrode surface of the molten drop-coated of Poly A, at 37 ℃ after incubation with its surface of TE damping fluid drip washing; Finally, by 5～20 μ mol L
-1quercetin-Cu ethanolic solution drip and be coated with its electrode surface, at 37 ℃ after incubation with its surface of TE damping fluid drip washing.
3. method according to claim 2, is characterized in that: (preparation method of 1) Zhong perylene tetracarboxylic acid/graphene oxide heterojunction is as follows: prepare 1 * 10 for described step
-4mol L
-1perylene tetracarboxylic acid DMF solution and 1mg mL
-1graphene oxide solution, Ran Hou perylene tetracarboxylic acid solution and graphene oxide solution with the volume ratio of 5:1 fully mix, standing, i.e. Sheng Cheng perylene tetracarboxylic acid/graphene oxide heterojunction.
4. method according to claim 2, is characterized in that: in described step (3), the activation method of modified electrode is as follows: by 20 μ L5mmol L
-1eDC and 10mmol L
-1the Tris-HCl solution Di of the pH7.4 of NHS is surperficial Zhi perylene tetracarboxylic acid/graphene oxide heterojunction modified electrode, and activates 1～2h under room temperature.
5. method according to claim 1, is characterized in that: by " On-Off " type aptamer based photoelectrochemicalcell sensor building in 0.1mol L
-1in phosphate buffered solution, at current potential 0.2V partially and be greater than the mercury ion that adopts electric current-time technology for detection variable concentrations under the optical radiation of 450nm.
6. method according to claim 5, is characterized in that: described method is hypersensitive and specific detection, detects and is limited to 3.3fmol L
-1.
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| CN104931570A (en) * | 2015-06-08 | 2015-09-23 | 济南大学 | Preparation method and application of electrochemical sensor for heavy metal ions based on aptamers |
| CN105004712A (en) * | 2015-07-17 | 2015-10-28 | 盐城工学院 | Method for constructing acetamiprid detection photoelectrochemical sensor and detecting method |
| CN105241938A (en) * | 2015-09-14 | 2016-01-13 | 盐城工学院 | Construction method and detection method of potassium ion nucleic acid aptamer photoelectrochemical sensor based on diluted magnetic semiconductor |
| CN105353006A (en) * | 2015-11-11 | 2016-02-24 | 安徽理工大学 | Photoelectric sensor and application thereof, and preparation method of working electrode |
| CN106841350A (en) * | 2017-02-07 | 2017-06-13 | 济南大学 | Electrochemical sensor based on aptamer detection mercury ion and preparation method thereof |
| CN108760852A (en) * | 2018-04-13 | 2018-11-06 | 江西师范大学 | Photoelectrochemical ochratoxin A detection method based on dual signal amplification |
| CN110687175A (en) * | 2019-11-11 | 2020-01-14 | 济南大学 | Construction method of electrochemical luminescence sensor based on cerium dioxide and nano-silver dual-enhanced perylene tetracarboxylic acid luminescence |
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| CN104459130A (en) * | 2014-09-26 | 2015-03-25 | 济南大学 | Aptamer-based biosensor for detecting penbritin and preparation method of aptamer-based biosensor |
| CN104931570A (en) * | 2015-06-08 | 2015-09-23 | 济南大学 | Preparation method and application of electrochemical sensor for heavy metal ions based on aptamers |
| CN104931570B (en) * | 2015-06-08 | 2016-04-20 | 济南大学 | A kind of preparation method of the heavy metal ion electrochemical sensor based on aptamer and application |
| CN105004712A (en) * | 2015-07-17 | 2015-10-28 | 盐城工学院 | Method for constructing acetamiprid detection photoelectrochemical sensor and detecting method |
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| CN105241938B (en) * | 2015-09-14 | 2017-11-17 | 盐城工学院 | A kind of structure and detection method of the potassium ion aptamer optical electro-chemistry sensor based on dilute magnetic semiconductor |
| CN105353006B (en) * | 2015-11-11 | 2018-05-15 | 安徽理工大学 | A kind of preparation method and application of photoelectric sensor and its working electrode |
| CN105353006A (en) * | 2015-11-11 | 2016-02-24 | 安徽理工大学 | Photoelectric sensor and application thereof, and preparation method of working electrode |
| CN106841350A (en) * | 2017-02-07 | 2017-06-13 | 济南大学 | Electrochemical sensor based on aptamer detection mercury ion and preparation method thereof |
| CN106841350B (en) * | 2017-02-07 | 2019-03-26 | 济南大学 | Electrochemical sensor and preparation method thereof based on aptamer detection mercury ion |
| CN108760852A (en) * | 2018-04-13 | 2018-11-06 | 江西师范大学 | Photoelectrochemical ochratoxin A detection method based on dual signal amplification |
| CN108760852B (en) * | 2018-04-13 | 2021-03-23 | 江西师范大学 | Photoelectrochemical ochratoxin A detection method based on dual signal amplification |
| CN110687175A (en) * | 2019-11-11 | 2020-01-14 | 济南大学 | Construction method of electrochemical luminescence sensor based on cerium dioxide and nano-silver dual-enhanced perylene tetracarboxylic acid luminescence |
| CN110687175B (en) * | 2019-11-11 | 2022-06-10 | 济南大学 | Construction method of electrochemical luminescence sensor based on cerium dioxide and nano-silver dual-enhanced perylene tetracarboxylic acid luminescence |
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