CN102879336B - Method for manufacturing plasma chiral ligand sensor for mercury ions - Google Patents
Method for manufacturing plasma chiral ligand sensor for mercury ions Download PDFInfo
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Abstract
The invention provides a method for manufacturing a plasma chiral ligand sensor for mercury ions, and belongs to the technical field of analytical chemistry and the technical field of food safety. The method mainly comprises the following steps of: (1) synthesizing gold nanorods; (2) performing directional nucleic acid functionalization on surfaces of the gold nanorods; (3) constructing the plasma chiral ligand sensor by using the gold nanorods after the surfaces of the gold nanorods are subjected to the directional nucleic acid functionalization; (4) representing the plasma chiral ligand sensor; (5) measuring the specificity of the plasma chiral ligand sensor; and (6) measuring an actual sample of the plasma chiral ligand sensor. Along with increase of Hg2+ concentration, the number of the gold nanorods in a gold nanorods side surface assembly body is increased, and the Mohr ellipticity of the assembly body is also increased. The linear relation between the Mohr ellipticity and the concentration of the mercury ions is set up, and the plasma chiral ligand sensor which is simple and sensitive and is high in specificity is manufactured and is used for detecting the concentration of the mercury ions.
Description
Technical field
A preparation method for the plasma chirality aptamers sensor of mercury ion, belongs to technical field of analytical chemistry and food security technical field.
Background technology
Nano material refers to that three dimensions yardstick has the material of one dimension in nanometer scale at least.Nano material is the material of new generation between cluster and macroscopic material.Because the size of nano material has approached the relevant wavelength of de Broglie wavelength, superconduction of electronics and the Bohr radius of exciton, electronics is limited to a space that volume is very small, electron wave function is restricted, add that it has the special effects on large surface, thereby there are quantum size effect, surface effect, small-size effect and macro quanta tunnel effect etc., so nano material demonstrates the character of novel optics, electricity, magnetics and the chemical aspect different from conventional body phase material.Will on the basis of single nano material character, there is new optics, electricity, magnetics and catalytic property in the assembling of nano material, (as: sensor, catalyzer, Magnetized Material and optical material etc.) have potential using value in a lot of fields, therefore, the research of nanoparticle assembly becomes and becomes increasingly active.
As everyone knows, circular dichroism refers to when linearly polarized light is when having the medium of optically active, because same optically active molecule in medium exists two kinds of configurations that chirality is different, therefore the dextrorotation that they resolve into linearly polarized light is different with Left-hand circular polarization light absorption, and the difference of absorption coefficient is defined as circular dichroism.Thereby circular dichroism spectrometer obtains the secondary structure of biomacromolecule by measuring the circular dichroism spectrum of biomacromolecule, be easy and obtain efficiently one of means of structure of biological macromolecule.Therefore, it can be used for the research of Dan white matter Zhe Die ﹑ protein conformation, DNA/RNA reaction, enzyme kinetics, optically active substance purity determination, Drugs.
In recent years, chirality nano material becomes the study hotspot problem that people pay close attention to, particularly, since within 2009, successfully building plasma nano particle chirality tetrahedron, this kind of chirality assembly impels scientific circles and industry member to give great concern for the discrete assembly of plasma nano material formation chirality.The chiral signal of plasma chirality assembly is different from the chiral signal of biomolecule, it not only has stronger signal at ultraviolet region (traditional biomolecule has and only have circular dichroism spectra signal in this region), and it has also produced extremely strong chiral signal in visible region.By research, find, this assembly is better than the signal of ultraviolet region at the productive rate signal closely bound up and its visible region of the intensity of the signal of visible region and the package assembly of plasma nano material, by foregoing description, can absolutely prove that plasma assembly can be successfully applied to bio-sensing detection field.Utilizing aptamer to carry out plasma nanomaterial assembly produces the circular dichroism spectra signal of visible region and then still belongs to blank for this field of detection of heavy metal ion.
The present invention utilizes base thymine (T) to carry out specific combination with mercury ion and forms T-Hg
2+-T structure, makes the gold nanorods single stranded DNA that coupling contains multiple thymine alkali bases (all the other sequence complete complementaries) respectively form plasma aptamers probe and forms the gold nanorods side assembling body with chiral signal, due to the Hg adding
2+concentration difference make the number of contained rod in side assembling body that gold nanorods forms different, and then cause the numerical value of molar ellipticity of gold nanorods side assembling body different (along with Hg
2+the increase of concentration, the number of the gold nanorods containing in gold nanorods side assembling body increases, the molar ellipticity of assembly is larger), set up the linear relationship of molar ellipticity and ion concentration of mercury, build the plasma chirality aptamers sensor of simple, sensitive and high specific.
Summary of the invention
The preparation method who the object of this invention is to provide a kind of plasma chirality aptamers sensor of mercury ion, has built the novel plasma chirality aptamers sensor of simple, the sensitive and high specific that utilizes circular dichroism spectra input.
Technical scheme of the present invention: a kind of preparation method of plasma chirality aptamers sensor of mercury ion, main implementation step of the present invention is: (1) gold nanorods synthetic; (2) gold nanorods surface orientation nucleic acid function; (3) utilize the gold nanorods of oriented nuclei acid functionalization to build plasma chirality aptamers sensor; (4) sign of plasma chirality aptamers sensor; (5) specific assay of plasma chirality aptamers sensor; (6) actual sample of plasma chirality aptamers sensor is measured.
Concrete steps are:
(1) gold nanorods is synthetic
The synthetic length-diameter ratio of method according to mono-kind of patent < < with the preparation method > > (patent No.: ZL 201010605799.9) of the self-assembled material of surface reinforced Raman active is about 3.0 gold nanorods.Simple step is as follows:
1. crystal seed is synthetic: the three hydration tetra chlorauric acids of getting the 2 pre-configured g/L of 0.1 mL at 28 ℃ join in the cetyl trimethyl ammonium bromide solution of 0.2 M of 1 mL, and now, solution colour is by the colourless tawny that becomes.Then to the 0.01 M sodium borohydride solution rapid stirring 2 minutes that adds the new preparation of 0.12 mL in above-mentioned mixed system.Solution colour becomes light brown from tawny.Above-mentioned synthetic crystal seed is put into the standing reaction of water-bath 30 minutes of 28 ℃, make not participate in completely the synthetic sodium borohydride approach exhaustion of crystal seed, (the one, the gold ion of complete reaction and being consumed not in the further reduction system of sodium borohydride, the 2nd, in standing course of reaction, sodium borohydride further reduces because hydrolysis causes its content), the crystal seed of preparation will carry out next step reaction in synthetic latter 4 hours.
2. gold nanorods growth course: after crystal seed has synthesized, carry out the growth of gold nanorods.The three hydration tetra chlorauric acids of 2 g/L of 5 mL add the ultrapure water of 4 mL after joining in the cetyl trimethyl ammonium bromide of 5 mL 0.2 M, mix.The 0.01 M liquor argenti nitratis ophthalmicus of 0.125 mL joins in above-mentioned mixed system, mixes, and subsequently the 0.1 M ascorbic acid solution of 65 μ L is added to above-mentioned system solution, and vigorous stirring reaction is 2 minutes at 28 ℃, and solution is become colourless by brown.Finally, add the seed-solution of 0.05 mL softly to stir and evenly mix 20 seconds.The color of 28 ℃ of reactions solution after 2 hours is drabon look.Synthetic gold nanorods solution is centrifugal 10 minutes through 10000 rpms, abandon supernatant, adopt the cetyl trimethyl ammonium bromide solution of 0.005 M that precipitation is heavily disperseed, clean once, the most at last gold nanorods Solution Dispersion with initial isopyknic solution in, the standing preservation of room temperature, to carry out next step sign and experiment demand.
(2) gold nanorods surface orientation nucleic acid function
1. gold nanorods end face sealing: the gold nanorods 1000 μ L that get above-mentioned 10 nM that prepare, polyglycol solution (molecular weight is 5000) the jolting reaction 8 hours of sulfydryl modification that adds 1 mM of 5 μ L, 7500 rpms centrifugal 15 minutes, abandon supernatant, washing once, with aqueous dispersion gold nanorods-polyethylene glycol complex of 1000 μ L, 4 ℃ of preservations, stand-by.
2. gold nanorods lateral orientation is modified APTa: the gold nanorods-polyglycol solution 500 μ L that get above-mentioned processing add the APTa of sulfydryl modification of the 100 μ M of 10 μ L, mix, room temperature jolting reaction 12 hours, 7500 rpms centrifugal 15 minutes, abandon supernatant, cetyl trimethyl ammonium bromide Solution Dispersion gold nanorods-polyglycol-APTa compound with 5 mM of 500 μ L, the cetyl trimethyl ammonium bromide solution of 5 mM is washed once, again use cetyl trimethyl ammonium bromide Solution Dispersion gold nanorods-polyglycol-APTa compound of 5 mM of 500 μ L, 4 ℃ of preservations, stand-by.
3. gold nanorods lateral orientation is modified APTb: the gold nanorods-polyglycol solution 500 μ L that get above-mentioned processing add the APTb of sulfydryl modification of the 100 μ M of 10 μ L, mix, room temperature jolting reaction 12 hours, 7500 rpms centrifugal 15 minutes, abandon supernatant, cetyl trimethyl ammonium bromide Solution Dispersion gold nanorods-polyglycol-APTb compound with 5 mM of 500 μ L, the cetyl trimethyl ammonium bromide solution of 5 mM is washed once, again use cetyl trimethyl ammonium bromide Solution Dispersion gold nanorods-polyglycol-APTb compound of 5 mM of 500 μ L, 4 ℃ of preservations, stand-by.
APTa：5’-SH-AAAAAAGTGACCATTTTTGCAGTG-?3’，
APTb：5’-SH-CACTGCTTTTTTGGTCACAAAAAA-3’。
(3) utilize the gold nanorods of oriented nuclei acid functionalization to build plasma chirality aptamers sensor
Circular dichroism spectra detects, draw molar ellipticity～ion concentration of mercury typical curve: get the gold nanorods that the APTb lateral orientation of gold nanorods that the APTa lateral orientation of the above-mentioned preparation of 50 μ L modifies and the above-mentioned preparation of 50 μ L modifies and mix, soft jolting reaction 5 minutes, the mercury ion standard items that add 50 μ L variable concentrations, hatch after 1 hour, by T-Hg for 40 ℃
2+the gold nanorods that-T specific recognition APTa and APTb modify respectively forms gold nanorods side assembling structure.Because the difference of the standard items concentration of the mercury ion adding causes the chiral signal generation difference that the length of the chain structure that gold nanorods forms is different and then shine into plasma chiral sensor.The standard items concentration of mercury ion is respectively 0 ng/mL, 0.05 ng/mL, 0.1 ng/mL, 0.5 ng/mL, 1 ng/mL, 5 ng/mL, 10ng/mL.Utilize the molar ellipticity of the package assembly under the different ion concentration of mercury of circular dichroism spectra Instrument measuring, according to the molar ellipticity of the variable concentrations of measuring under 730nm wavelength, draw the typical curve of ion concentration of mercury.The range of linearity of this sensor is 0.05 to 10ng/mL, detection is limited to 0.03ng/mL, the linear equation of typical curve is that Y=0.5X-19.75(Y is the molar ellipticity under 730nm under variable concentrations, the concentration that X is corresponding mercury ion), linear dependence is >0.99.
(4) sign of plasma chirality aptamers sensor
Under mercury ion variable concentrations condition, the chirality aptamers sensor that adopts the instruments such as transmission electron microscope (accelerating potential is 200 keV), X-ray energy spectrometer, dynamic laser light scattering experimental instrument to build gold nanorods self assembly characterizes.By known this kind of chirality aptamers sensor of above-mentioned characterization method, successfully build.
(5) specific assay of plasma chirality aptamers sensor
According to the gold nanorods that utilizes oriented nuclei acid functionalization, build the same steps of in plasma chirality aptamers sensor step, mercury ion being measured and measure the 4 heavy metal species ion (Cu that likely exist in other actual sample
2+, Cr
6+, Fe
2+and Zn
2+), concentration is 10 ng/mL, by measuring the molar ellipticity of other heavy metal ion in above-mentioned 4, with the Hg of 10 ng/mL
2+the molar ellipticity of the plasma chirality aptamers sensor being shone is more known, and this kind of plasma chirality aptamers sensor tip is to mercury ion detecting high specificity.
(6) actual sample of plasma chirality aptamers sensor is measured
Using tap water as actual sample, verify that this kind of method is for the interpolation recovery of mercury ion.In advance to the mercury ion that adds 0.1 ng/mL, 0.5 ng/mL, 1 ng/mL and 5 ng/mL in tap water sample used, adopt the interpolation recovery of the mercury ion in plasma chirality aptamers sensor mensuration actual sample between 94.81-103.10%, standard deviation is 3.5%, can meet the detection demand to mercury ion in actual sample completely.
Beneficial effect of the present invention: along with Hg
2+the increase of concentration, the number of the gold nanorods containing in gold nanorods side assembling body increases, the molar ellipticity of assembly is larger, the present invention has set up the linear relationship of molar ellipticity and ion concentration of mercury, build the plasma chirality aptamers sensor of the mercury ion of simple, sensitive and high specific, for detection of the concentration of mercury ion.
Accompanying drawing explanation
Ultraviolet/visible light spectrogram before and after Fig. 1 gold nanorods coupling aptamers probe APTa and APTb
The transmission electron microscope picture of plasma chirality aptamers sensor under the typical different ion concentration of mercury conditions of Fig. 2, (a-g) concentration of mercury ion is respectively 0,0.05,0.1,0.5,1,5 and 10 ng/mL and (h) the low power transmission electron microscope picture under 5ng/mL concentration.
The X-ray energy spectrum figure of Fig. 3 plasma chirality aptamers sensor
The particle diameter of Fig. 4 plasma chirality aptamers sensor (a), rear (b) before adding the reaction of 10ng/mL mercury ion distributes
The circular dichroism spectrogram of Fig. 5 plasma chirality aptamers sensor under different ion concentration of mercury conditions, along with the direction that arrow refers to, the concentration of mercury ion increases gradually according to 0,0.05,0.1,0.5,1,5 and 10 ng/mL gradients.
Fig. 6 under 733nm, the typical curve of mercury ion plasma chirality aptamers sensor
The specificity result of Fig. 7 plasma chirality aptamers sensor.
Embodiment
(1) gold nanorods is synthetic
The synthetic length-diameter ratio of method according to mono-kind of patent < < with the preparation method > > (patent No.: ZL 201010605799.9) of the self-assembled material of surface reinforced Raman active is about 3.0 gold nanorods.Simple step is as follows:
1. crystal seed is synthetic: the three hydration tetra chlorauric acids of getting the 2 pre-configured g/L of 0.1 mL at 28 ℃ join in the cetyl trimethyl ammonium bromide solution of 0.2 M of 1 mL, and now, solution colour is by the colourless tawny that becomes.Then to the 0.01 M sodium borohydride solution rapid stirring 2min that adds the new preparation of 0.12 mL in above-mentioned mixed system.Solution colour becomes light brown from tawny.Above-mentioned synthetic crystal seed is put into standing reaction 30 min of water-bath of 28 ℃, make not participate in completely the synthetic sodium borohydride approach exhaustion of crystal seed, (the one, the gold ion of complete reaction and being consumed not in the further reduction system of sodium borohydride, the 2nd, in standing course of reaction, sodium borohydride further reduces because hydrolysis causes its content), the crystal seed of preparation will carry out next step reaction in 4h after synthetic.
2. gold nanorods growth course: after crystal seed has synthesized, carry out the growth of gold nanorods.The three hydration tetra chlorauric acids of 2 g/L of 5 mL add the ultrapure water of 4 mL after joining in the cetyl trimethyl ammonium bromide of 5 mL 0.2 M, mix.The 0.01 M liquor argenti nitratis ophthalmicus of 0.125 mL joins in above-mentioned mixed system, mixes, and subsequently the 0.1 M ascorbic acid solution of 65 μ L is added to above-mentioned system solution, and vigorous stirring is reacted 2 min at 28 ℃, and solution is become colourless by brown.Finally, add the seed-solution of 0.05 mL softly to stir and evenly mix 20s.After 28 ℃ of reaction 2 h, the color of solution is drabon look.Synthetic gold nanorods solution is through 10000 rpms of centrifugal 10 min, abandon supernatant, adopt the cetyl trimethyl ammonium bromide solution of 0.005 M that precipitation is heavily disperseed, clean once, the most at last gold nanorods Solution Dispersion with initial isopyknic solution in, the standing preservation of room temperature, to carry out next step sign and experiment demand.
(2) gold nanorods surface orientation nucleic acid function
1. gold nanorods end face sealing: the gold nanorods 1000 μ L that get above-mentioned 10 nM that prepare, polyglycol solution (molecular weight is 5000) the jolting reaction 8h of sulfydryl modification that adds 1 mM of 5 μ L, centrifugal 15 min of 7500 r/min, abandon supernatant, aqueous dispersion gold nanorods-polyethylene glycol complex with 1000 μ L, wash once 4 ℃ of preservations, stand-by.
2. gold nanorods lateral orientation is modified APTa: the gold nanorods-polyglycol solution 500 μ L that get above-mentioned processing add the APTa of sulfydryl modification of the 100 μ M of 10 μ L, mix, room temperature jolting reaction 12h, the centrifugal 15min of 7500 r/min, abandon supernatant, cetyl trimethyl ammonium bromide Solution Dispersion gold nanorods-polyglycol-APTa compound with 5 mM of 500 μ L, the cetyl trimethyl ammonium bromide solution of 5 mM is washed once, again use cetyl trimethyl ammonium bromide Solution Dispersion gold nanorods-polyglycol-APTa compound of 5 mM of 500 μ L, 4 ℃ of preservations, stand-by.
3. gold nanorods lateral orientation is modified APTb: the gold nanorods-polyglycol solution 500 μ L that get above-mentioned processing add the APTb of sulfydryl modification of the 100 μ M of 10 μ L, mix, room temperature jolting reaction 12h, centrifugal 15 minutes of 7500 r/min, abandon supernatant, cetyl trimethyl ammonium bromide Solution Dispersion gold nanorods-polyglycol-APTb compound with 5 mM of 500 μ L, the cetyl trimethyl ammonium bromide solution of 5 mM is washed once, again use cetyl trimethyl ammonium bromide Solution Dispersion gold nanorods-polyglycol-APTb compound of 5 mM of 500 μ L, 4 ℃ of preservations, stand-by.
APTa：5’-SH-AAAAAAGTGACCATTTTTGCAGTG-?3’，
APTb：5’-SH-CACTGCTTTTTTGGTCACAAAAAA-3’。
(3) utilize the gold nanorods of oriented nuclei acid functionalization to build plasma chirality aptamers sensor
Circular dichroism spectra detects, draw molar ellipticity～ion concentration of mercury typical curve: get the gold nanorods that the APTb lateral orientation of gold nanorods that the APTa lateral orientation of the above-mentioned preparation of 50 μ L modifies and the above-mentioned preparation of 50 μ L modifies and mix, soft jolting reaction 5min, the mercury ion standard items that add 50 μ L variable concentrations, hatch after 1h for 40 ℃, by T-Hg
2+the gold nanorods that-T specific recognition APTa and APTb modify respectively forms gold nanorods side assembling structure.Because the difference of the standard items concentration of the mercury ion adding causes the chiral signal generation difference that the length of the chain structure that gold nanorods forms is different and then shine into plasma chiral sensor.The standard items concentration of mercury ion is respectively 0,0.05,0.1,0.5,1,5,10ng/mL.Utilize the molar ellipticity of the package assembly under the different ion concentration of mercury of circular dichroism spectra Instrument measuring, according to the molar ellipticity of the variable concentrations of measuring under 730nm wavelength, draw the typical curve of ion concentration of mercury.The range of linearity of this sensor is 0.05 to 10ng/mL, detection is limited to 0.03ng/mL, the linear equation of typical curve is that Y=0.5X-19.75(Y is the molar ellipticity under 730nm under variable concentrations, the concentration that X is corresponding mercury ion), linear dependence is >0.99.
(4) sign of plasma chirality aptamers sensor
Under mercury ion variable concentrations condition, the chirality aptamers sensor that adopts the instruments such as transmission electron microscope (accelerating potential is 200 keV), X-ray energy spectrometer, dynamic laser light scattering experimental instrument to build gold nanorods self assembly characterizes.By known this kind of chirality aptamers sensor of above-mentioned characterization method, successfully build.
(5) specific assay of plasma chirality aptamers sensor
According to the gold nanorods that utilizes oriented nuclei acid functionalization, build the same steps of in plasma chirality aptamers sensor step, mercury ion being measured and measure the 4 heavy metal species ion (Cu that likely exist in other actual sample
2+, Cr
6+, Fe
2+and Zn
2+), concentration is 10 ng/mL, by measuring the molar ellipticity of other heavy metal ion in above-mentioned 4, with the Hg of 10 ng/mL
2+the molar ellipticity of the plasma chirality aptamers sensor being shone is known, and this kind of plasma chirality aptamers sensor tip is to mercury ion detecting high specificity.
(6) actual sample of plasma chirality aptamers sensor is measured
Using tap water as actual sample, verify that this kind of method is for the interpolation recovery of mercury ion.In advance to the mercury ion that adds 0.1 ng/mL, 0.5 ng/mL, 1 ng/mL and 5 ng/mL in tap water sample used, adopt the interpolation recovery of the mercury ion in plasma chirality aptamers sensor mensuration actual sample between 94.81-103.10%, standard deviation is 3.5%, can meet the detection demand to mercury ion in actual sample completely.
The interpolation recovery test result of table 1 plasma chirality aptamers sensor
A: in TAIHU LAKE, the original concentration of mercury ion is to adopt atomic fluorescence spectrometer to measure
B: standard deviation is to calculate by measuring the concentration of the mercury ion of three samples under each concentration conditions.
Claims (1)
1. a preparation method for the plasma chirality aptamers sensor of mercury ion, is characterized in that gold nanorods surface orientation nucleic acid function, utilizes gold nanorods structure plasma chirality aptamers sensor, the sign of plasma chirality aptamers sensor, the actual sample of the specific assay of plasma chirality aptamers sensor and plasma chirality aptamers sensor of oriented nuclei acid functionalization to measure; Technique is
(1) gold nanorods surface orientation nucleic acid function
1. gold nanorods end face sealing: the gold nanorods 1000 μ L that get 10 nM that prepare, the polyglycol solution that the molecular weight of sulfydryl modification that adds 1 mM of 5 μ L is 5000, jolting reaction 8h, the centrifugal 15min of 7500r/min, abandon supernatant, wash once, with aqueous dispersion gold nanorods-polyethylene glycol complex of 1000 μ L, 4 ℃ of preservations, stand-by;
2. gold nanorods lateral orientation is modified APTa: get 1. gold nanorods-polyglycol solution 500 μ L of gained of step, the APTa of sulfydryl modification that adds the 100 μ M of 10 μ L, mix, room temperature jolting reaction 12h, the centrifugal 15min of 7500r/min, abandon supernatant, cetyl trimethyl ammonium bromide Solution Dispersion gold nanorods-polyglycol-APTa compound with 5 mM of 500 μ L, the cetyl trimethyl ammonium bromide solution of 5 mM is washed once, again use cetyl trimethyl ammonium bromide Solution Dispersion gold nanorods-polyglycol-APTa compound of 5 mM of 500 μ L, 4 ℃ of preservations, stand-by,
3. gold nanorods lateral orientation is modified APTb: get 1. gold nanorods-polyglycol solution 500 μ L of gained of step, the APTb of sulfydryl modification that adds the 100 μ M of 10 μ L, mix, room temperature jolting reaction 12h, the centrifugal 15min of 7500r/min, abandon supernatant, cetyl trimethyl ammonium bromide Solution Dispersion gold nanorods-polyglycol-APTb compound with 5 mM of 500 μ L, the cetyl trimethyl ammonium bromide solution of 5 mM is washed once, again use cetyl trimethyl ammonium bromide Solution Dispersion gold nanorods-polyglycol-APTb compound of 5 mM of 500 μ L, 4 ℃ of preservations, stand-by,
APTa：5’-SH-AAAAAAGTGACCATTTTTGCAGTG-?3’，
APTb：5’-SH-CACTGCTTTTTTGGTCACAAAAAA-3’，
(2) utilize the gold nanorods of oriented nuclei acid functionalization to build plasma chirality aptamers sensor
Circular dichroism spectra detects, draw molar ellipticity～ion concentration of mercury typical curve: get the gold nanorods that the APTb lateral orientation of gold nanorods that the APTa lateral orientation of the above-mentioned preparation of 50 μ L modifies and the above-mentioned preparation of 50 μ L modifies and mix, soft jolting reaction 5min, the mercury ion standard items that add 50 μ L variable concentrations, hatch after 1h for 40 ℃, by T-Hg
2+the gold nanorods that-T specific recognition APTa and APTb modify respectively forms gold nanorods side assembling structure; Because the difference of the standard items concentration of the mercury ion adding causes the length of the chain structure that gold nanorods forms different and then cause the chiral signal of plasma chiral sensor to produce difference; The standard items concentration of mercury ion is respectively 0 ng/mL, 0.05 ng/mL, 0.1 ng/mL, 0.5 ng/mL, 1 ng/mL, 5 ng/mL, 10ng/mL; Utilize the molar ellipticity of the package assembly under the different ion concentration of mercury of circular dichroism spectra Instrument measuring, according to the molar ellipticity of the variable concentrations of measuring under 730nm wavelength, draw the typical curve of ion concentration of mercury; The range of linearity of this sensor is 0.05-10ng/mL, detection is limited to 0.03ng/mL, and the linear equation of typical curve is Y=0.5X-19.75, and Y is the molar ellipticity under 730nm under variable concentrations, X is the concentration of corresponding mercury ion, and linear dependence is >0.99;
(3) sign of plasma chirality aptamers sensor
Under mercury ion variable concentrations condition, employing accelerating potential is that the chirality aptamers sensor that transmission electron microscope, X-ray energy spectrometer, the dynamic laser light scattering experimental instrument of 200 keV builds gold nanorods self assembly characterizes, and by known this kind of chirality aptamers sensor of above-mentioned characterization method, successfully builds;
(4) specific assay of plasma chirality aptamers sensor
According to the gold nanorods that utilizes oriented nuclei acid functionalization, build the same steps of in plasma chirality aptamers sensor step, mercury ion being measured and measure the 4 heavy metal species ion Cu that likely exist in other actual sample
2+, Cr
6+, Fe
2+and/or Zn
2+, concentration is 10 ng/mL, by measuring one or more molar ellipticity of above-mentioned 4 heavy metal species ions, with the Hg of 10 ng/mL
2+the molar ellipticity of the plasma chirality aptamers sensor causing is more known, and this kind of plasma chirality aptamers sensor tip is to mercury ion detecting high specificity;
(5) actual sample of plasma chirality aptamers sensor is measured
Using tap water as actual sample, verify that this kind of method is for the interpolation recovery of mercury ion, in advance to the mercury ion that adds respectively 0.1 ng/mL, 0.5 ng/mL, 1 ng/mL and 5 ng/mL in tap water sample used, adopt the interpolation recovery of the mercury ion in plasma chirality aptamers sensor mensuration actual sample between 94.81%-103.10%, standard deviation is 3.5%, can meet the detection demand to mercury ion in actual sample completely.
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| CN102590170B (en) * | 2012-02-28 | 2013-11-20 | 江南大学 | Method for simultaneously detecting mercury ion and/or silver ion in water solution based on fluorescence resonance energy transfer |
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