CN105675519A - Mercury ion detection method - Google Patents
Mercury ion detection method Download PDFInfo
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- CN105675519A CN105675519A CN201610202599.6A CN201610202599A CN105675519A CN 105675519 A CN105675519 A CN 105675519A CN 201610202599 A CN201610202599 A CN 201610202599A CN 105675519 A CN105675519 A CN 105675519A
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- BQPIGGFYSBELGY-UHFFFAOYSA-N mercury(2+) Chemical compound [Hg+2] BQPIGGFYSBELGY-UHFFFAOYSA-N 0.000 title claims abstract description 34
- 238000001514 detection method Methods 0.000 title claims abstract description 23
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims abstract description 42
- 238000006243 chemical reaction Methods 0.000 claims abstract description 39
- 239000001509 sodium citrate Substances 0.000 claims abstract description 21
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims abstract description 21
- 229920001213 Polysorbate 20 Polymers 0.000 claims abstract description 16
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 claims abstract description 16
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 claims abstract description 16
- 230000031700 light absorption Effects 0.000 claims abstract description 15
- 238000002156 mixing Methods 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 103
- 238000000034 method Methods 0.000 claims description 26
- 239000000872 buffer Substances 0.000 claims description 20
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 claims description 12
- 238000010521 absorption reaction Methods 0.000 claims description 10
- 102000039446 nucleic acids Human genes 0.000 claims description 10
- 108020004707 nucleic acids Proteins 0.000 claims description 10
- 150000007523 nucleic acids Chemical class 0.000 claims description 10
- 239000012279 sodium borohydride Substances 0.000 claims description 7
- 229910000033 sodium borohydride Inorganic materials 0.000 claims description 7
- 229910001961 silver nitrate Inorganic materials 0.000 claims description 6
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims description 5
- 229910052700 potassium Inorganic materials 0.000 claims description 5
- 239000011591 potassium Substances 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 4
- 239000002245 particle Substances 0.000 claims description 3
- 229910052697 platinum Inorganic materials 0.000 claims description 3
- 229910052709 silver Inorganic materials 0.000 claims description 3
- 102000003992 Peroxidases Human genes 0.000 abstract description 5
- 108040007629 peroxidase activity proteins Proteins 0.000 abstract description 5
- 230000035945 sensitivity Effects 0.000 abstract description 3
- 235000013305 food Nutrition 0.000 abstract description 2
- 229910052751 metal Inorganic materials 0.000 abstract description 2
- 239000002184 metal Substances 0.000 abstract description 2
- 230000003139 buffering effect Effects 0.000 abstract 1
- 229910052753 mercury Inorganic materials 0.000 abstract 1
- -1 mercury ions Chemical class 0.000 abstract 1
- 238000006555 catalytic reaction Methods 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- 239000008399 tap water Substances 0.000 description 4
- 229910021642 ultra pure water Inorganic materials 0.000 description 4
- 239000012498 ultrapure water Substances 0.000 description 4
- 230000003197 catalytic effect Effects 0.000 description 3
- 239000002086 nanomaterial Substances 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 102000053602 DNA Human genes 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- 238000005374 membrane filtration Methods 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 235000020679 tap water Nutrition 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- SZVJSHCCFOBDDC-UHFFFAOYSA-N ferrosoferric oxide Chemical compound O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000001965 increasing effect Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- Physics & Mathematics (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a mercury ion detection method. The mercury ion detection method comprises the following steps: adding a solution to be detected, which possibly contains Hg<2+>, Tween-20, a sodium citrate buffering solution, hydrogen peroxide and a TBM solution, into a double-metal nano cluster solution, and mixing for reacting; and detecting the concentration of Hg<2+> in the solution to be detected by observing a light absorption value of a visible light wave band of a mixing reaction system. The mercury ions (Hg<2+>) are detected colorimetrically by simulating peroxidase based on DNA-Ag/Pt nano clusters; a linear range of detection is 10.0nM-200nM and the sensitivity can reach 3.0nM; and the mercury ion detection method has the advantages of simplicity, convenience and rapidness, low cost, high stability and the like, and can be applied to detection of Hg<2+> in samples of environments, foods and the like.
Description
Technical field
The present invention relates to the detection method of a kind of mercury ion, utilize single-chain nucleic acid to prepare bimetal nano bunch the method that it is applied to mercury ion detecting particularly to a kind of, belong to nanosecond science and technology field.
Background technology
Enzyme is protein or the nucleic acid that a class has catalysis, it is possible to catalytic chemistry reaction in suitable environment. But, enzyme is biocatalyzer, and due to the characteristic that enzyme is intrinsic, required for it, the condition of catalysis is relatively stricter, and needs higher cost, and its application has been also affected by some restrictions. The easy degeneration of such as protease and hydrolysis, relatively costly and use condition strict. Therefore, a kind of analogue enztme that there is similar catalysis active is developed particularly important.
Nanometer analogue enztme is a class non-protein structure but has the synthetic catalyst of similar catalytic performance to native enzyme; except there is catalytic stability height, preparing the advantages such as easy, cheap, production scale; can also at room temperature preserve stable; preparation, purification and storage are all easier; it is prone to modify and labelling types of functionality molecule or the advantage of protein antibodies, causes the extensive concern of research worker.
Since the report ferroferric oxide nano granules such as Institute of Biophysics, Academia Sinica's usury increasings in 2007 have peroxidase original mold plan activity, substantial amounts of nano material is in the news as analogue enztme and has peroxidase activity. Wherein, introducing the second metal in monometallic nano material all can change to overall catalysis activity and selectivity, and therefore bimetal nano material is increasingly becoming the focus that people pay close attention to. Therefore, exploitation nanometer analogue enztme has significantly high application prospect.
Mercury ion is widely present in the several samples such as environment, food, due to it many biologies are had high toxicity and there is cumulative toxicity, thus, how a kind of quick detection that nanometer analogue enztme is applied to mercury ion is provided, also becomes the direction of industry research staff research.
Summary of the invention
For above-mentioned deficiency of the prior art, it is an object of the invention to provide a kind of mercury ion detecting method, its can fast and easy, low cost, highly sensitive, high stability realize Hg2+Colorimetric detection.
For realizing aforementioned invention purpose, the technical solution used in the present invention includes:
A kind of mercury ion detecting method, including: add in bimetal nano cluster solution and be likely to containing Hg2+Solution to be measured, sodium citrate buffer, hydrogen peroxide and TMB solution hybrid reaction, by observing the hybrid reaction system light absorption value at visible light wave range, it is achieved to the Hg in solution to be measured2+The detection of concentration.
Further, the method includes:
The standard Hg of tween 20, sodium citrate buffer, hydrogen peroxide, TMB solution and a series of variable concentrations is added in DNA-Ag/Pt nanocluster solution2+Solution hybrid reaction, measures the light absorption value at visible light wave range, it is thus achieved that Hg2+Concentration-light absorption value standard curve;
Sodium citrate buffer, hydrogen peroxide, TMB solution and solution to be measured, hybrid reaction is added in DNA-Ag/Pt nanocluster solution, thus the Hg recorded in solution to be measured2+Concentration.
One of preferably, described DNA-Ag/Pt nanocluster particle diameter is at 3.0nm～6.0nm.
One of preferably, in described DNA-Ag/Pt nanocluster, the mass ratio of Ag and Pt is 1:10～1:30.
One of preferably, the concentration of described DNA-Ag/Pt nanocluster solution is 0.1 μM～2.0 μMs.
One of preferably, described tween 20 concentration is 0.005%-0.05%.
One of preferably, the pH value of described sodium citrate buffer is 3.0～4.5.
One of preferably, the concentration of described hydrogen peroxide is 0.5M～2.0M.
One of preferably, the concentration of described TMB (TMB) solution is 1.0mM～4.0mM.
One of preferably, described reaction temperature is at 20 DEG C～45 DEG C.
Further, the method comprises the steps:
(1) the DNA-Ag/Pt nanocluster solution providing concentration to be 0.1 μM～2.0 μMs;
(2) taking 8 μ L step (1) the DNA-Ag/Pt nanocluster solution obtained, the sodium citrate buffer, the 20 μ L concentration that are sequentially added into 2 μ L concentration to be the tween 20 of 0.005%-0.05%, 30 μ LpH values be 3.0～4.5 are the standard Hg of the hydrogen peroxide of 0.5M～2.0M and TMB solution that 40 μ L concentration are 1.0mM～4.0mM and the 100 a series of variable concentrations of μ L2+Solution, is uniformly mixed to form hybrid reaction system, after reacting 10min when 20 DEG C～45 DEG C, then measures each hybrid reaction system light absorption value at visible light wave range respectively, and obtains Hg2+Concentration-light absorption value standard curve;
(3) 8 μ L step (1) the DNA-Ag/Pt nanocluster solution obtained is taken, and to be sequentially added into 2 μ L concentration be the tween 20 of 0.005%-0.05%, 30 μ LpH values are the sodium citrate buffer of 3.0～4.5, 20 μ L concentration are the hydrogen peroxide of 0.5M～2.0M and TMB solution that 40 μ L concentration are 1.0mM～4.0mM and 100 μ L solution to be measured, it is uniformly mixed to form hybrid reaction system, after reacting 10min when 20 DEG C～45 DEG C, measure this hybrid reaction system light absorption value at visible light wave range again, and with described standard curve control, thus the Hg recorded in solution to be measured2+Concentration.
Concrete, the preparation method of described DNA-Ag/Pt nanocluster solution includes (list of references chemicalcommunications2014,50 (86), 13103-6.):
After silver nitrate solution that the single-chain nucleic acid solution that concentration is 2.0 μMs that volume ratio is 30:5:12, concentration are 150 μMs and Tetrachloroplatinate potassium solution Homogeneous phase mixing that concentration is 125 μMs, adding concentration after lucifuge reaction 30min at 4 DEG C is the sodium borohydride solution of 5.0mM, the volume ratio of described single-chain nucleic acid solution and sodium borohydride solution is 30:4, vibrate at 37 DEG C 3h, obtains the DNA-Ag/Pt nanocluster solution that concentration is 2.0 μMs.
Wherein, the sequence of the typical single-chain nucleic acid of employing is: 5 '-CCCCCTAACTCCCCC-3 ', but is not limited to this.
Compared with prior art, advantages of the present invention at least that: the present invention is based on DNA-Ag/Pt nanocluster Mimetic enzyme colorimetric detection mercury ion (Hg2+), the range of linearity of its detection is 10.0-200nM, and sensitivity can reach 3.0nM; Have easy quickly, cost low, stability advantages of higher, can be applicable to Hg in the sample such as environment, food2+Detection.
Accompanying drawing explanation
Fig. 1 is Hg in the present invention one typical embodiments2+Suppress DNA-Ag/Pt nanocluster solution (DNA-Ag/PtNCs) Mimetic Peroxidase catalysis active principle figure;
Fig. 2 is at Hg in one embodiment of the invention2+It is absent from (a) and there is the absorption collection of illustrative plates of DNA-Ag/Pt nanocluster solution Mimetic Peroxidase catalysis activity time (b);
Fig. 3 is obtained Hg in the embodiment of the present invention 12+Concentration-light absorption value standard curve;
Fig. 4 is the selectivity test collection of illustrative plates in the embodiment of the present invention 1 for different metal ion.
Detailed description of the invention
For making the object, technical solutions and advantages of the present invention clearly, below in conjunction with accompanying drawing, the specific embodiment of the present invention is described in detail. The example of these preferred implementations has illustrated in the accompanying drawings. Shown in accompanying drawing and the embodiments of the present invention that describe with reference to the accompanying drawings merely exemplary, and the present invention is not limited to these embodiments.
At this, in addition it is also necessary to explanation, in order to avoid having obscured the present invention because of unnecessary details, illustrate only in the accompanying drawings and according to the closely-related structure of the solution of the present invention and/or process step, and eliminate other details little with relation of the present invention.
A kind of mercury ion detecting method, including: add in bimetal nano cluster solution and be likely to containing Hg2+Solution to be measured and sodium citrate buffer, hydrogen peroxide and TMB solution hybrid reaction, by observing the hybrid reaction system light absorption value at visible light wave range, it is achieved to the Hg in solution to be measured2+The detection of concentration.
Further, the method includes:
Tween 20, sodium citrate buffer, hydrogen peroxide, TMB solution and standard Hg is added in DNA-Ag/Pt nanocluster solution2+Solution hybrid reaction, measures the light absorption value at visible light wave range, it is thus achieved that Hg2+Concentration-light absorption value standard curve;
Tween 20, sodium citrate buffer, hydrogen peroxide, TMB solution and solution to be measured, hybrid reaction is added in DNA-Ag/Pt nanocluster solution, thus the Hg recorded in solution to be measured2+Concentration.
One of preferably, described DNA-Ag/Pt nanocluster particle diameter is at 3.0nm～6.0nm.
One of preferably, in described DNA-Ag/Pt nanocluster, the mass ratio of Ag and Pt is 1:10～1:30.
One of preferably, the concentration of described DNA-Ag/Pt nanocluster solution is 0.1 μM～2.0 μMs.
One of preferably, the concentration of described tween 20 is 0.005%-0.05%.
One of preferably, the pH value of described sodium citrate buffer is 3.0～4.5.
One of preferably, the concentration of described hydrogen peroxide is 0.5M～2.0M.
One of preferably, the concentration of described TMB (TMB) solution is 1.0mM～4.0mM.
One of preferably, described reaction temperature is at 20 DEG C～45 DEG C.
Further, the method comprises the steps:
(1) the DNA-Ag/Pt nanocluster solution providing concentration to be 0.1 μM～2.0 μMs;
(2) taking 8 μ L step (1) the DNA-Ag/Pt nanocluster solution obtained, the sodium citrate buffer, the 20 μ L concentration that are sequentially added into 2 μ L concentration to be the tween 20 of 0.005%-0.05%, 30 μ LpH values be 3.0～4.5 are the standard Hg of the hydrogen peroxide of 0.5M～2.0M and TMB solution that 40 μ L concentration are 1.0mM～4.0mM and the 100 a series of variable concentrations of μ L2+Solution, is uniformly mixed to form hybrid reaction system, after reacting 10min when 20 DEG C～45 DEG C, then measures each hybrid reaction system light absorption value at visible light wave range respectively, and obtains Hg2+Concentration-light absorption value standard curve;
(3) 8 μ L step (1) the DNA-Ag/Pt nanocluster solution obtained is taken, and to be sequentially added into 2 μ L concentration be the tween 20 of 0.005%-0.05%, 30 μ LpH values are the sodium citrate buffer of 3.0～4.5, 20 μ L concentration are the hydrogen peroxide of 0.5M～2.0M and TMB solution that 40 μ L concentration are 1.0mM～4.0mM and 100 μ L solution to be measured, it is uniformly mixed to form hybrid reaction system, after reacting 10min when 20 DEG C～45 DEG C, measure this hybrid reaction system light absorption value at visible light wave range again, and with described standard curve control, thus the Hg recorded in solution to be measured2+Concentration.
Concrete, the preparation method of described DNA-Ag/Pt nanocluster solution includes:
The PCR pipe of 200 μ L is sequentially added into 30 μ L, the single-chain nucleic acid (ssDNA) of 2.0 μMs, 5 μ L, the silver nitrate solution of 150 μMs and 12 μ L, the Tetrachloroplatinate potassium solution of 125 μMs, adds the sodium borohydride solution of 4 μ L, 5.0mM after lucifuge reaction 30min at 4 DEG C, vibrate at 37 DEG C 3h, obtains the DNA-Ag/Pt nanocluster solution of 2.0 μMs.
In example 1 below-2, involved raw material, as silver nitrate, Platinous Potassium Chloride, ssDNA (sequence as indicated earlier, but can be not limited to this) and other reagent all can be obtained by commercially available approach.
Embodiment 1
(1) according to the list of references synthetic DNA-Ag/PtNCs reported, specifically comprise the following steps that
The PCR pipe of 200 μ L is sequentially added into 30 μ L, the single-chain nucleic acid of 2.0 μMs, 5 μ L, the silver nitrate solution of 150 μMs and 12 μ L, the Tetrachloroplatinate potassium solution of 125 μMs, adds the sodium borohydride solution of 4 μ L, 5.0mM after lucifuge reaction 30min at 4 DEG C, vibrate at 37 DEG C 3h, obtains the DNA-Ag/Pt nanocluster solution of 2.0 μMs.
(2) taking ultra-pure water and dilute final the obtained mixed solution 8 μ L of step (1) of 2 times, the sodium citrate buffer, the 20 μ L concentration that are sequentially added into 2 μ L concentration to be the tween 20 of 0.005%-0.05%, 30 μ LpH values be 3.0～4.5 are the standard Hg of the hydrogen peroxide of 0.5M～2.0M and TMB solution that 40 μ L concentration are 1.0mM～4.0mM and the 100 a series of variable concentrations of μ L2+Solution, is uniformly mixed to form hybrid reaction system, after reacting 10min when 20 DEG C～45 DEG C, then measures each hybrid reaction system light absorption value at visible light wave range respectively, and obtains Hg2+Concentration-light absorption value standard curve, as it is shown on figure 3, detection sensitivity can reach 3.0nM, the detection range of linearity is 10-200nM.
(3) Hg in tap water2+Detection: by tap water through 0.22 μm of microporous filter membrane filtration, ultra-pure water dilutes 3 times. With reference to above-mentioned steps (2) method, replace Hg with the tap water sample after simple process2+Solution, can complete Hg in tap water2+Detection.
It addition, the operation of reference step (2) and essentially identical reaction condition, other metal ion also having carried out the detection of comparison, its final detection result see Fig. 4.
Embodiment 2
(1) according to the list of references synthetic DNA-Ag/PtNCs reported, specifically comprise the following steps that
The PCR pipe of 200 μ L is sequentially added into 30 μ L, the single-chain nucleic acid of 2.0 μMs (is referred to embodiment 1, but it is not limited to this), 5 μ L, the silver nitrate solution of 150 μMs and 12 μ L, the Tetrachloroplatinate potassium solution of 125 μMs, 4 μ L are added after lucifuge reaction 30min at 4 DEG C, the sodium borohydride solution of 5.0mM, vibrate at 37 DEG C 3h, obtains the DNA-Ag/Pt nanocluster solution of 2.0 μMs.
(2) taking ultra-pure water and dilute final the obtained mixed solution 8 μ L of step (1) of 4 times, the sodium citrate buffer, the 20 μ L concentration that are sequentially added into 2 μ L concentration to be the tween 20 of 0.005%-0.05%, 30 μ LpH values be 3.0～4.5 are the standard Hg of the hydrogen peroxide of 0.5M～2.0M and TMB solution that 40 μ L concentration are 1.0mM～4.0mM and the 100 a series of variable concentrations of μ L2+Solution, is uniformly mixed to form hybrid reaction system, measures each hybrid reaction system light absorption value at visible light wave range more respectively, and obtain ion concentration of mercury-light absorption value standard curve after reacting 10min when 20 DEG C～45 DEG C.
(3) Hg in river2+Detection: by river through 0.22 μm of microporous filter membrane filtration, ultra-pure water dilutes 10 times. With reference to above-mentioned steps (2) method, replace Hg with the river sample after simple process2+Solution, can complete Hg in river2+Detection.
It is pointed out that above-described embodiment is only the technology design and feature that the present invention is described, its object is to allow person skilled in the art will appreciate that present disclosure and to implement according to this, can not limit the scope of the invention with this. All equivalences made according to spirit of the invention change or modify, and all should be encompassed within protection scope of the present invention.
Claims (10)
1. a mercury ion detecting method, it is characterised in that including: add in bimetal nano cluster solution and be likely to containing Hg2+Solution to be measured, sodium citrate buffer, hydrogen peroxide and TMB solution hybrid reaction, by observing the hybrid reaction system light absorption value at visible light wave range, it is achieved to the Hg in solution to be measured2+The detection of concentration; Described bimetal nano cluster is DNA-Ag/Pt nanocluster.
2. mercury ion detecting method according to claim 1, it is characterised in that the method includes:
The standard Hg of sodium citrate buffer, hydrogen peroxide, TMB solution and a series of variable concentrations is added in DNA-Ag/Pt nanocluster solution2+Solution hybrid reaction, measures the light absorption value at visible light wave range, it is thus achieved that Hg2+Concentration-light absorption value standard curve;
Tween 20, sodium citrate buffer, hydrogen peroxide, TMB solution and solution to be measured, hybrid reaction is added in DNA-Ag/Pt nanocluster solution, thus the Hg recorded in solution to be measured2+Concentration.
3. mercury ion detecting method according to claim 2, it is characterised in that described DNA-Ag/Pt nanocluster particle diameter is at 3.0nm～6.0nm.
4. mercury ion detecting method according to claim 2, it is characterised in that in described DNA-Ag/Pt nanocluster, the mass ratio of Ag and Pt is 1:10～1:30.
5. mercury ion detecting method according to claim 2, it is characterised in that the concentration of described DNA-Ag/Pt nanocluster solution is 0.1 μM～2.0 μMs.
6. mercury ion detecting method according to claim 2, it is characterised in that the concentration of described tween 20 is 0.005wt%-0.05wt%.
7. mercury ion detecting method according to claim 2, it is characterised in that the pH value of described sodium citrate buffer is 3.0～4.5; And/or, the concentration of described hydrogen peroxide is 0.5M～2.0M; And/or, the concentration of described TMB solution is 1.0mM～4.0mM.
8. mercury ion detecting method according to claim 2, it is characterised in that: described reaction temperature is at 20 DEG C～45 DEG C.
9. the mercury ion detecting method according to any one of claim 1-8, it is characterised in that the method comprises the steps:
(1) the DNA-Ag/Pt nanocluster solution providing concentration to be 0.1 μM～2.0 μMs;
(2) taking 8 μ L step (1) the DNA-Ag/Pt nanocluster solution obtained, the sodium citrate buffer, the 20 μ L concentration that are sequentially added into 2 μ L concentration to be the tween 20 of 0.005%-0.05%, 30 μ LpH values be 3.0～4.5 are the standard Hg of the hydrogen peroxide of 0.5M～2.0M and TMB solution that 40 μ L concentration are 1.0mM～4.0mM and the 100 a series of variable concentrations of μ L2+Solution, is uniformly mixed to form hybrid reaction system, after reacting 10min when 20 DEG C～45 DEG C, then measures each hybrid reaction system light absorption value at visible light wave range respectively, and obtains Hg2+Concentration-light absorption value standard curve;
(3) 8 μ L step (1) the DNA-Ag/Pt nanocluster solution obtained is taken, and to be sequentially added into 2 μ L concentration be the tween 20 of 0.005%-0.05%, 30 μ LpH values are the sodium citrate buffer of 3.0～4.5, 20 μ L concentration are the hydrogen peroxide of 0.5M～2.0M and TMB solution that 40 μ L concentration are 1.0mM～4.0mM and 100 μ L solution to be measured, it is uniformly mixed to form hybrid reaction system, after reacting 10min when 20 DEG C～45 DEG C, measure this hybrid reaction system light absorption value at visible light wave range again, and with described standard curve control, thus the Hg recorded in solution to be measured2+Concentration.
10. mercury ion detecting method according to claim 9, it is characterized in that, the preparation method of described DNA-Ag/Pt nanocluster solution includes: by the single-chain nucleic acid solution that concentration is 2.0 μMs that volume ratio is 30:5:12, concentration is after the silver nitrate solution of 150 μMs and Tetrachloroplatinate potassium solution Homogeneous phase mixing that concentration is 125 μMs, lucifuge reaction 30min at 4 DEG C, adding concentration afterwards is the sodium borohydride solution of 5.0mM, the volume ratio of described single-chain nucleic acid solution and sodium borohydride solution is 30:4, vibrate 3h again at 37 DEG C, obtain the DNA-Ag/Pt nanocluster solution that concentration is 2.0 μMs.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520972A (en) * | 2016-11-25 | 2017-03-22 | 复旦大学 | Method for detecting nucleic acid concentration by using self-assembly system based on nucleic acid-platinum nano-material |
| CN106546585A (en) * | 2016-11-09 | 2017-03-29 | 江南大学 | Mercury ion, total mercury and organomercurial detection method and detection kit |
| CN107356578A (en) * | 2017-08-16 | 2017-11-17 | 广西师范大学 | A kind of aptamers regulate and control silica nanometer enzymatic activity resonance scattering spectroscopy measure Hg2+Method |
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| CN106546585A (en) * | 2016-11-09 | 2017-03-29 | 江南大学 | Mercury ion, total mercury and organomercurial detection method and detection kit |
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| CN107807117A (en) * | 2017-08-16 | 2018-03-16 | 广西师范大学 | A kind of aptamers regulate and control silica nanometer enzymatic activity SERS measure Hg2+Method |
| CN107807117B (en) * | 2017-08-16 | 2019-09-24 | 广西师范大学 | It is a kind of to measure Hg with aptamers regulation silica nanometer enzymatic activity Surface enhanced Raman spectroscopy2+Method |
| CN109030473A (en) * | 2018-06-13 | 2018-12-18 | 盐城工学院 | Utilize the method for Nano silver grain detection mercury ion |
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| CN114807146A (en) * | 2022-06-28 | 2022-07-29 | 中国农业大学 | Preparation method and sensing application of nucleic acid nanoenzyme |
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