CN106111974A - A kind of preparation method and application of gold silver core-shell particles gold nanorods self-assembled structures - Google Patents
A kind of preparation method and application of gold silver core-shell particles gold nanorods self-assembled structures Download PDFInfo
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- CN106111974A CN106111974A CN201610592438.2A CN201610592438A CN106111974A CN 106111974 A CN106111974 A CN 106111974A CN 201610592438 A CN201610592438 A CN 201610592438A CN 106111974 A CN106111974 A CN 106111974A
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Abstract
The preparation method and application of a kind of gold silver core-shell particles gold nanorods self-assembled structures, belongs to analytical chemistry field.The present invention includes the synthesis of gold silver core-shell nano, the synthesis of gold nanorods, the nucleic acid of gold silver core-shell nano is modified, the fluorescent dye of gold nanorods and aptamer modified, the dimeric self assembly of gold silver core-shell particles gold nanorods, Raman beacon is modified, and the bimodal detection by quantitative of dopamine.This self-assembled structures integrates surface enhanced raman spectroscopy (SERS) and fluorescence signal, the gold silver core-shell particles gold nanorods dimer having surface enhanced raman spectroscopy and fluorescence signal concurrently of formation；Along with change and the surface plasma resonance character of regulation and control package assembly of gold nanorods and the spacing of gold silver core-shell particles, by monitoring the change of its SERS and fluorescence signal intensity, dopamine can be carried out the most quantitatively.
Description
Technical field
The present invention relates to the preparation method and application of a kind of gold silver core-shell particles-gold nanorods self-assembled structures, belong to point
Analysis chemical field.
Background technology
Since nano material is born, due to the quantum size effect of its excellence, skin effect, quantum tunneling effect, inhale
Draw the extensive concern of numerous scientist and research workers, science and society's every field have been had deep effect.Base
In the cooperative effect of assembling primitive, the assembly of nano material not only has the characteristic of nano-particle, the most also can produce new
Coupling effect, shows the character such as optics, electricity and magnetics of uniqueness, and become analytical chemistry, optics, catalysis, chemical industry,
The ideal material of the numerous areas such as environment and medical science.Dopamine (DA) is a kind of important neurotransmitters, the change of its content
May result in senile dementia and schizophrenia etc., therefore the research of its assay method is to pathogenic mechanism studies and clinical application
There is important practical significance.The detection method of dopamine at present, including HPLC method, ultraviolet visible spectrophotometry, electrification
, high performance capillary electrophoresis, chemoluminescence method etc..Although these quantitative detecting methods have certain sensitivity and specificity, but
It is all single modal response signal, it is impossible to guarantee precisely to analyze.
The present invention is based on gold silver core-shell particles-gold nanorods self-assembled structures, it is achieved that SERS and the bimodulus of fluorescence signal
The hypersensitive detection by quantitative of the dopamine of state.
Summary of the invention
It is an object of the invention to provide the preparation method of a kind of gold silver core-shell particles-gold nanorods self-assembled structures and answer
With, the most do not prepare the report of gold silver core-shell particles-gold nanorods self-assembled structures.The self-assembled structures of the present invention, with
Unlike other structures existing, modify dyefunctionalized aptamer at the end face of gold nanorods, and nucleic acid is modified
Gold silver core-shell particles carry out self assembly, be simultaneously introduced Raman beacon, ultimately form and have surface enhanced raman spectroscopy and fluorescence concurrently
Gold silver core-shell particles-gold nanorods the dimer of signal.
The method that the present invention also aims to simultaneously provide a kind of bimodal detection dopamine, say, that along with Jenner
The change of the spacing of rice rod and gold silver core-shell particles and the surface plasma resonance character of regulation and control package assembly, by monitoring it
SERS and the change of fluorescence signal intensity, can be carried out dopamine the most quantitatively.
Technical scheme, the preparation method of a kind of gold silver core-shell particles-gold nanorods self-assembled structures, including such as
The step of lower order:
(1) synthesis of golden nanometer particle: take and add 97.5mL ultra-pure water in the round-bottomed flask of cleaning, add the 4g/L's of 2.5mL
Chlorauric acid solution, temperature constant magnetic stirring mixes and is heated to boiling, and keeps seething with excitement after 5-6min, fast in above-mentioned mixed solution
Speed adds the citric acid three sodium solution of the 1% of 2.0mL, and continuous heating stirring 10min, until solution is stable bright claret-red
Color, finally, reaction solution is cooled to room temperature, and 4 DEG C of preservations are standby, i.e. prepares the golden nanometer particle that particle diameter is 18nm；
(2) synthesis of gold silver core-shell particles: first the golden nanometer particle of 100 μ L of pre-synthesis, is centrifuged and is resuspended in containing 0.5%
In the phosphate buffered solution of the 5mM of polyvinylpyrrolidone (PVP), afterwards, the silver nitrate of the 2.75mM adding 30 μ L is molten
Liquid, is simultaneously introduced the ascorbic acid of 0.1M of 20 μ L as reducing agent, and stirring reaction 3h under room temperature, centrifugation, by precipitate
After cleaning three times with ultra-pure water, being resuspended in the ultra-pure water of 100 μ L, 4 DEG C of preservations are standby, i.e. obtain gold silver coreshell type structure nanometer
Material；
(3) synthesis of gold nanorods:
1. crystal seed synthesis: the chlorauric acid solution of the 4g/L taking 0.1mL in clean conical flask joins 1mL while stirring
0.2M cetyl trimethylammonium bromide (CTAB) solution in, solution colour is become yellowish-brown by colourless；It is subsequently adding
The 0.01M sodium borohydride solution that 0.12mL newly prepares quickly stirs 2min, and solution colour is i.e. become light brown from yellowish-brown；
2. gold nanorods growth: in the CTAB solution of the 0.2M that the 4g/L chlorauric acid solution taking 5mL joins 5mL, and add 4mL
Ultra-pure water；The 0.1M ascorbic acid solution of the 0.01M silver nitrate solution and 65 L that separately take 0.125mL is added separately to above-mentioned mixed
In fit system, stirring reaction 2min at 28 DEG C, solution is become colourless by brown；It is eventually adding 0.05mL step 1. gained crystal seed molten
30 DEG C of standing and reacting 3h after liquid stirring 20s, obtain gold nanorods solution；
(4) nucleic acid of gold silver core-shell particles is modified: takes the 100 μ L gold silver core-shell particles 10000rpm that step (2) synthesizes and is centrifuged
10min, after removing supernatant, precipitation is resuspended in the 10mM PB solution of 100 μ L, adds DNA1, and molar concentration is than gold silver nucleocapsid
Particle DNA1 be 12 coupling ratio carry out coupling, stand 12h, centrifugal, be resuspended in 100 μ L 10mM PB solution, obtain Au@
Ag-DNA1 complex；
(5) nucleic acid of gold nanorods is modified: takes the scattered 10mL gold nanorods 7000rpm that step (3) synthesizes and is centrifuged 10min,
After removing supernatant, precipitation is resuspended in 10mL 5mM CTAB solution, adds 5 ' end sulfydryl modifications, the DOPA of 3 ' end Cy5 dyestuffs modifications
Amine aptamer DNA2, and molar concentration carries out coupling than the coupling ratio that gold nanorods DNA2 is 12, stands 12h, from
The heart, is resuspended in 10mL 10mM PB solution, obtains GNR-DNA2 complex；
(6) gold silver core-shell particles-gold nanorods self-assembly: Au@Ag-DNA1 that step (4) and step (5) are obtained and
The complex of GNR-DNA2 respectively takes 100 μ L and carries out equal-volume mixing, stands 12h, adds afterwards and make final concentration reach 15 μMs
Raman beacon molecule ATP, i.e. can get SERS and the fluorescence signal gold silver core-shell particles-gold nanorods dimer in one from group
Assembling structure；
Described DNA1:5 '-GGG CCT CAT TCT GTG CGA ACG CTT TTG TAC CGC ACA GCC TCT GGC
GCA CAC AGA GAC -3′；
DNA2 i.e. DA-aptamer:5 '-SH-GTC TCT GTG TGC GCC AGA GAC ACT GGG GCA GAT ATG
GGC CAG CAC AGA ATG AGG CCC-Cy5 -3′。
(7) the bimodal quantitative determination of dopamine: Au@Ag-DNA1 and GNR-that step (4) and step (5) are obtained
The complex of DNA2 respectively takes 100 μ L and carries out equal-volume mixing, is simultaneously introduced 5 μ L dopamine to be measured sample solution, stands 12h,
Add the Raman beacon molecule ATP making final concentration reach 15 μMs afterwards, measure the Raman letter of dopamine sample system the most respectively
Number and fluorescence signal, thus, the concentration of dopamine can be detected by the situation of change of Raman signal and fluorescence signal.
The quantitative dopamine concentration of bimodal；Under fluorescence mode, the Monitoring lower-cut of dopamine concentration is 0.05fM, Raman modes
The Monitoring lower-cut of lower dopamine concentration is 0.03fM.
Beneficial effects of the present invention: the present invention modifies dyefunctionalized aptamer at the end face of gold nanorods,
The gold silver core-shell particles modified with nucleic acid carries out self assembly, is simultaneously introduced Raman beacon, ultimately forms and have surface-enhanced Raman concurrently
Scattering and the gold silver core-shell particles-gold nanorods dimer of fluorescence signal；Spacing along with gold nanorods Yu gold silver core-shell particles
Change and regulation and control package assembly surface plasma resonance character, by monitoring the change of its SERS and fluorescence signal intensity,
Dopamine can be carried out the most quantitatively.
Accompanying drawing explanation
The transmission electron microscope picture of Fig. 1 gold silver core-shell particles-gold nanorods assembly.
The Raman collection of illustrative plates of Fig. 2 quantitative dopamine based on gold silver core-shell particles-gold nanorods assembly.
The fluorescence pattern of Fig. 3 quantitative dopamine based on gold silver core-shell particles-gold nanorods assembly.
Detailed description of the invention
Embodiment 1
All of glass apparatus all soaks 24h with chloroazotic acid, and cleans with distilled water, dries standby.The water used in experiment is
18.2 the Milli-Q ultra-pure water of M Ω.
(1) synthesis of golden nanometer particle: take and add 97.5mL ultra-pure water in the round-bottomed flask of cleaning, add 2.5mL's
The chlorauric acid solution of 4g/ L, temperature constant magnetic stirring mixes and is heated to boiling, and keeps seething with excitement after 5-6min, to above-mentioned mixing
Solution is rapidly added 2.0mL 1% citric acid three sodium solution, continuous heating stirring 10min, until solution is stable saturating
Bright claret.Finally, reaction solution is cooled to room temperature, and 4 DEG C of preservations are standby, can prepare the gold that particle diameter is about 18 nm
Nanoparticle.
(2) synthesis of gold silver core-shell particles: first the golden nanometer particle of 100 μ L of pre-synthesis, centrifugal be resuspended in containing
In the phosphate buffered solution of the 5mM of 0.5% PVP.Afterwards, add the silver nitrate solution of the 2.75mM of 30 μ L, be simultaneously introduced 20 μ
The ascorbic acid of the 0.1M of L is as reducing agent, and stirring reaction 3h under room temperature, centrifugation, by precipitate ultra-pure water cleaning three
After secondary, being resuspended in the ultra-pure water of 100 μ L, 4 DEG C of preservations are standby, i.e. can get gold silver nano material coreshell type structure.
(3) synthesis of gold nanorods:
1. crystal seed synthesis: the chlorauric acid solution taking the 4g/L taking 0.1mL in the conical flask of cleaning joins 1mL's while stirring
In cetyl trimethylammonium bromide (CTAB) solution of 0.2M, solution colour is become yellowish-brown by colourless；It is subsequently adding
The 0.01M sodium borohydride solution that 0.12mL newly prepares quickly stirs 2min, and solution colour is i.e. become light brown from yellowish-brown.
2. gold nanorods growth: in the CTAB solution of the 0.2M that the 4g/L chlorauric acid solution taking 5mL joins 5mL, and add
Enter 4mL ultra-pure water；The 0.1M ascorbic acid solution of the 0.01M silver nitrate solution and 65 L that separately take 0.125mL is added separately to
Stating in mixed system, stirring reaction 2min at 28 DEG C, solution is become colourless by brown；It is eventually adding 0.05mL step 1. gained
30 DEG C of standing and reacting 3h after seed-solution stirring 20s, obtain gold nanorods solution.
(4) nucleic acid of gold silver core-shell particles is modified: take 100 μ L gold silver core-shell particles 10000rpm that step (2) synthesizes from
Heart 10min, after removing supernatant, precipitation is resuspended in the 10mM PB solution of 100 μ L, adds DNA1, and molar concentration is than gold silver core
Shell particles DNA1 be 12 coupling ratio carry out coupling, stand 12h, centrifugal, be resuspended in 100 μ L 10mM PB solution, obtain
Au@Ag-DNA1 complex；
(5) nucleic acid of gold nanorods is modified: takes the scattered 10mL gold nanorods 7000rpm that step (3) synthesizes and is centrifuged 10min,
After removing supernatant, precipitation is resuspended in 10mL 5mM CTAB solution, adds 5 ' end sulfydryl modifications, the DOPA of 3 ' end Cy5 dyestuffs modifications
Amine aptamer DNA2, and molar concentration carries out coupling than the coupling ratio that gold nanorods DNA2 is 12, stands 12h,
Centrifugal, it is resuspended in 10mL 10mM PB solution, obtains GNR-DNA2 complex.
DNA1:5 '-SH-GGG CCT CAT TCT GTG CGA ACG CTT TTG TAC CGC ACA GCC TCT
GGC GCA CAC AGA GAC -3′；
DNA2 (DA-aptamer): 5 '-GTC TCT GTG TGC GCC AGA GAC ACT GGG GCA GAT ATG GGC
CAG CAC AGA ATG AGG CCC-Cy5 -3′。
(6) gold silver core-shell particles-gold nanorods self-assembly: the Au@Ag-DNA1 that step (4) and step (5) are obtained
And the complex of GNR-DNA2 respectively takes 100 μ L and carries out equal-volume mixing, stand 12h, add afterwards and make final concentration reach 15 μMs
Raman beacon molecule ATP, i.e. can get SERS and fluorescence signal in one gold silver core-shell particles-gold nanorods dimer.
The transmission electron microscope picture of gold silver core-shell particles-gold nanorods assembly is as shown in Figure 1.
(7) the bimodal quantitative determination of dopamine: Au@Ag-DNA1 and GNR-that step (4) and step (5) are obtained
The complex of DNA2 respectively takes 100 μ L and carries out equal-volume mixing, is simultaneously introduced 5 μ L dopamine to be measured sample solution, stands 12h,
Add the Raman beacon molecule ATP making final concentration reach 15 μMs afterwards, measure the Raman letter of dopamine sample system the most respectively
Number and fluorescence signal, thus, the concentration of dopamine can be detected by the situation of change of Raman signal and fluorescence signal.
The Raman collection of illustrative plates of quantitative dopamine based on gold silver core-shell particles-gold nanorods assembly is as shown in Figure 2；Based on gold
The fluorescence pattern of the quantitative dopamine of galactic nucleus shell particles-gold nanorods assembly is as shown in Figure 3.
Claims (4)
1. the preparation method of gold silver core-shell particles-gold nanorods self-assembled structures, it is characterised in that step is as follows:
(1) nucleic acid of gold silver core-shell particles is modified: takes the gold silver core-shell particles that 100 μ L prepare and is centrifuged with 10000 rpm
10min, after removing supernatant, precipitation is resuspended in the 10mM PB solution of 100 μ L, adds DNA1, and molar concentration is than gold silver core
Shell particles DNA1 be 12 coupling ratio carry out coupling, stand 12h, centrifugal, be resuspended in 100 μ L 10mM PB solution, obtain
Au@Ag-DNA1 complex；
(2) nucleic acid of gold nanorods is modified: takes the scattered 10mL gold nanorods prepared and is centrifuged 10min with 7000rpm, goes
After supernatant, precipitation is resuspended in the 5mM CTAB solution of 10mL, adds 5 ' end sulfydryl modifications, the DOPA of 3 ' end Cy5 dyestuffs modifications
Amine aptamer DNA2, and molar concentration carries out coupling than the coupling ratio that gold nanorods DNA2 is 12, stands 12h, from
The heart is resuspended in 10mL 10mM PB solution, obtains GNR-DNA2 complex；
(3) gold silver core-shell particles-gold nanorods self-assembly: Au@Ag-DNA1 step (1) prepared and step (2) prepare
GNR-DNA2 complex respectively takes 100 μ L and carries out equal-volume mixing, stands 12h, adds afterwards and makes final concentration reach 15 μMs draw
Graceful beacon molecule ATP, i.e. obtains SERS and fluorescence signal in the gold silver core-shell particles-gold nanorods self-assembled structures of one.
The preparation method of gold silver core-shell particles-gold nanorods self-assembled structures the most according to claim 1, it is characterised in that:
Described DNA1:5 '-GGG CCT CAT TCT GTG CGA ACG CTT TTG TAC CGC ACA GCC TCT GGC GCA
CAC AGA GAC -3′；
DNA2 i.e. DA-aptamer:5 '-SH-GTC TCT GTG TGC GCC AGA GAC ACT GGG GCA GAT ATG
GGC CAG CAC AGA ATG AGG CCC-Cy5 -3′。
3. the application of gold silver core-shell particles-gold nanorods self-assembled structures that prepared by method described in claim 1, its feature exists
In: it is applied to dopamine detection, specifically comprises the following steps that what Au@Ag-DNA1 step (1) prepared and step (2) prepared
GNR-DNA2 complex respectively takes 100 μ L and carries out equal-volume mixing, is simultaneously introduced 5 μ L dopamine to be measured sample solution, stands
12h, adds the Raman beacon molecule ATP making final concentration reach 15 μMs afterwards, measures drawing of dopamine sample system the most respectively
Graceful signal and fluorescence signal, thus, detected the concentration of dopamine by the situation of change of Raman signal and fluorescence signal.
The application of gold silver core-shell particles-gold nanorods self-assembled structures the most according to claim 3, it is characterised in that: bimodulus
The quantitative dopamine concentration of state；Under fluorescence mode, the Monitoring lower-cut of dopamine concentration is 0.05fM, dopamine concentration under Raman modes
Monitoring lower-cut be 0.03fM.
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| CN108070826A (en) * | 2017-11-20 | 2018-05-25 | 西北工业大学 | A kind of method for preparing gold electrode surfaces dopamine self-assembled monolayer |
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| CN111204705A (en) * | 2020-01-14 | 2020-05-29 | 大连理工大学 | Preparation method of directionally assembled core-cap heterogeneous dimer structure |
| CN111204705B (en) * | 2020-01-14 | 2021-05-07 | 大连理工大学 | Preparation method of directionally assembled core-cap heterogeneous dimer structure |
| CN113567413A (en) * | 2021-06-18 | 2021-10-29 | 南京大学 | Method for detecting monoamine neurotransmitters based on low-frequency Raman scattering technology |
| CN114047172A (en) * | 2021-11-04 | 2022-02-15 | 北京大学 | Method for quenching biological background fluorescence to realize Raman spectrum detection |
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