CN107418561B - Blue-fluorescence gold nano point, preparation method and its application in bivalent cupric ion context of detection - Google Patents
Blue-fluorescence gold nano point, preparation method and its application in bivalent cupric ion context of detection Download PDFInfo
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Abstract
A kind of blue-fluorescence gold nano point, preparation method and its application in bivalent cupric ion context of detection, belong to fluorogold technical field of nano material.It is that NaOH aqueous solution is added in deionized water, adds polyfunctional group macromolecular aqueous solution, is then added and contains Au3+Aqueous solution to get arrive large-sized gold nanoparticle aqueous solution；NaOH aqueous solution is added thereto, the biomolecule aqueous solution containing sulfydryl collects supernatant, and by supernatant repeated centrifugation 3~5 times, finally obtains nanodot aqueous solution.The synthetic method is simple, mild condition, it is reproducible, it is easy to mass production, product chemistry property is stablized, fluorescence quantum efficiency is high, has overdelicate detection for bivalent cupric ion, detection is limited down to 0.7nM, and linear relationship is presented between the fluorescence intensity of concentration and the gold nano point of bivalent cupric ion, quantitative detection can be thus carried out to bivalent cupric ion, and there is very high selectivity, the almost influence of interference-free ion.
Description
Technical field
The invention belongs to fluorogold technical field of nano material, and in particular to a kind of blue-fluorescence gold nano point, preparation side
Method and its application in bivalent cupric ion context of detection.
Background technique
Noble metal nano point, due to having optics, electricity, the chemical property etc. that are totally different from conventional metals nano dot,
Causing extensive research interest (Chem.Soc.Rev., 2012,41,3594-3623) in recent years.Wherein gold nano point due to
Have many advantages, such as easily prepared, excellent photoluminescent property and chemical stability, is widely used in fluorescence probe, fluorescence sense etc.
Field (Microchimica Acta., 2014,182,695-701).
Bivalent cupric ion is the essential trace elements of the human body, but unsuitable intake copper ion, can be caused instead serious
Body illness.When intake copper ion is excessive, metabolic disturbance, hepato-biliary function obstacle etc. can be caused.Take in content of copper ion mistake
It is few, it will cause anaemia, osteoporosis, or even influence the brain growth etc. of infant.Thus, the detection of copper ion has been caused
Extensive research (J.Colloid Interface Sci., 2013,396,63-68).
Since conventional probe synthesis process is complicated, limit for height is detected, the reasons such as detection time length limit application.So anxious
It need to develop that a kind of synthesis process is simple, detects time-consuming short, the fluorescence probe of high sensitivity.
Summary of the invention
The object of the present invention is to provide a kind of blue-fluorescence gold nano point, preparation method and its in bivalent cupric ion detection side
The application in face.The method comprises the steps of firstly, preparing the gold nanoparticles of bulky grain, then have blue-fluorescence using the method preparation of etching
Gold nano point, and then copper ion is sensitively detected.
The gold nano spot size of preparation is uniform, diameter be less than 3nm, in the way of fluorescent quenching can with quantitative detection copper from
Son, detection limit is down to 0.7nM, and the detection method has very high selectivity for copper ion.
The preparation method of nanodot of the present invention and its as follows to the detecting step of copper ion:
1) in 20~30mL deionized water, being separately added into 0.25~1mL concentration is 0.01~1mol/L (preferably 0.1
~1mol/L) NaOH aqueous solution, addition concentration be 0.001~1mol/L (preferably 0.001~0.1mol/L, further it is excellent
Be selected as 0.001~0.01mol/L) polyfunctional group macromolecular aqueous solution (can be tetramethylol chloride, bovine serum albumin(BSA)
Deng aqueous solution) be used as stabilizer and reducing agent, wherein the dosage molar ratio of NaOH and polyfunctional group macromolecular be 0.1~10:
1；Then solution is stirred to 5~10min under 20~30 DEG C (preferably 25~30 DEG C), then rapidly join concentration be 1~
100mmol/L (preferably 1~50mmol/L, further preferably 1~10mmol/L) contains Au3+Aqueous solution (can be
HAuCl4、AuCl3Deng aqueous solution), polyfunctional group macromolecular and Au3+Dosage molar ratio be 10~1000:1；Solution is existed
Under 20~30 DEG C (preferably 25~30 DEG C) stir 20~60min (preferably 20~40min, further preferably 20~
30min) to get large-sized gold nanoparticle aqueous solution is arrived, it is stored in 4 DEG C, spare, the concentration of gold nanoparticle at this time
For 0.5~10mmol/L；
2) be in the gold nanoparticle aqueous solution for preparing of 0.5~10mmol/L in concentration be added concentration be 1~
The NaOH aqueous solution of 100mmol/L (preferably 1~50mmol/L), adding concentration is 0.01~1mol/L (preferably 0.01
~0.5mol/L, further preferably 0.01~0.05mol/L) biomolecule aqueous solution (cysteine, paddy Guang containing sulfydryl
Sweet peptide, the aqueous solution containing mercaptoamino acid, albumen containing sulfydryl etc.) it is used as stabilizer and etching agent；Wherein gold nanoparticle is water-soluble
The dosage molar ratio of liquid and the biomolecule containing sulfydryl is 0.01~0.1:1, the dosage molar ratio of NaOH and the biomolecule containing sulfydryl
For 0.1~1:1；Solution is stirred under 80~150 DEG C (preferably 100~150 DEG C, further preferably 100~120 DEG C) again
20~30h (preferably 20~25h), collects supernatant in eccentric fashion after reaction, and by supernatant repeated centrifugation 3~
5 times, finally obtained supernatant is nanodot aqueous solution of the present invention；
3) the nanodot aqueous solution 0.5mL for taking step 2) to prepare, be added 0.5mL concentration be respectively 0,2nM,
10nM, 1 μM, 50 μM, 100 μM, 150 μM, 200 μM, 300 μM, 500 μM, the aqueous solution containing bivalent cupric ion of 1mM, 2.5mM
(it can be CuCl2、CuSO4、Cu(NO3)2Deng aqueous solution), add 2mL deionized water solution, above-mentioned solution be put into glimmering
Its fluorescence intensity is tested in photothermal spectroscopic analyzer.Its fluorescence spectra shows, with the increase that copper ion concentration is added, gold nano point
Fluorescence intensity is gradually reduced, and copper ion concentration detection limit is calculated down to 0.7nM, and between copper ion concentration and fluorescence intensity
Good linear relationship is presented.
4) the nanodot aqueous solution 0.5mL for taking step 2) to prepare is firstly added the survey of 2.5mL deionized water solution
Its fluorescence intensity is tried, value is set as I0；It is then respectively adding 0.5mL and Cu2+Isoconcentration contains K+Aqueous solution (can be
KCl、K2SO4、KNO3Deng aqueous solution), Na+Aqueous solution (can be NaCl, Na2SO4、NaNO3Deng aqueous solution), Mg2+'s
Aqueous solution (can be MgCl2、MgSO4、Mg(NO3)2Deng aqueous solution), Zn2+Aqueous solution (can be ZnCl2、ZnSO4、Zn
(NO3)2Deng aqueous solution), Fe3+Aqueous solution (can be FeCl3、Fe2(SO4)3、Fe(NO3)3Deng aqueous solution), add
2mL deionized water；Above-mentioned solution is put into Fluorescence Spectrometer and tests its fluorescence intensity, value is set as I.Draw (I0- I)/I0
Column diagram shows that nanodot to the Difference test of different metal ions, shows nanodot prepared by the present invention
Selective enumeration method can be carried out to bivalent cupric ion.
Nanodot prepared by the present invention is for having the following characteristics that this to the super sensitivity detection of bivalent cupric ion
The synthetic method of nanodot is simple, and mild condition is reproducible, is easy to mass production, product chemistry property is stablized, glimmering
Photo-quantum efficiency is high, has overdelicate detection for bivalent cupric ion, detection limit is down to 0.7nM, and bivalent cupric ion is dense
Linear relationship is presented between degree and the fluorescence intensity of gold nano point, thus quantitative detection can be carried out to bivalent cupric ion, and should
Detection method shows very high selectivity for bivalent cupric ion, almost the influence of interference-free ion.
Detailed description of the invention
Fig. 1: Fig. 1 a is the TEM figure of nanodot prepared by embodiment 1, and diameter is about 1.9nm；Fig. 1 b is real
The TEM figure of nanodot prepared by example 3 is applied, diameter is about 2.5nm；Prepared gold nano point is circle, point
It dissipates uniformly, size is uniform.
Fig. 2: detection curve of the nanodot prepared by embodiment 1 for bivalent cupric ion.From different copper ions
Under concentration gold nano point fluorogram (Fig. 2 a) it can be seen that, be added copper ion concentration increase, gold nano point it is glimmering
Luminous intensity is gradually reduced, and detection is limited down to 0.7nM.It can from copper ion concentration and maximum fluorescence intensity numerical value mapping (Fig. 2 b)
It arrives, good linear relationship is presented between copper ion concentration and maximum fluorescence intensity.
Fig. 3: selective enumeration method figure of the nanodot prepared by embodiment 1 for bivalent cupric ion.In gold nano
It is separately added into and Cu in point aqueous solution2+The K of isoconcentration equivalent+、Na+、Mg2+、Zn2+、Fe3+, find Cu2+It can be by gold nano point
Fluorescence intensity be quenched 60% or more, and other impurities ion has little effect the fluorescence intensity of gold nano point.Show
The detection method has very high selectivity for copper ion.Wherein I0It is strong for gold nanoclusters fluorescence that metal ion is not added
Degree, I are the fluorescence intensity of gold nanoclusters after respective metal ion is added.
Specific embodiment
Embodiment 1
The NaOH aqueous solution that 0.25mL concentration is 1mol/L is separately added into 22.5mL deionized water, 3 μ L concentration are
The tetramethylol chloride aqueous solution of 10mmol/L, stirs 5min at 30 DEG C, and it is 50mmol/L's that 500 μ L concentration, which are then added,
HAuCl4Aqueous solution stirs 25min at 25 DEG C, and gold nanoparticle aqueous solution can be obtained, and obtains gold nanoparticle water at this time
Solution concentration is 1mmol/L.
30 μ L concentration are added as the NaOH aqueous solution of 1mol/L in the gold nanoparticle aqueous solution for taking 10mL to prepare, then plus
Enter 0.0138g cysteine, is stirred for 24 hours at 100 DEG C.Collect supernatant in eccentric fashion after reaction, and by supernatant
Liquid repeated centrifugation 3 times, finally obtaining supernatant is nanodot aqueous solution of the present invention.Under transmission electron microscope, gold
Nano-dot size is 1.9nm or so, uniform particle sizes, good dispersion (Fig. 1 a).
The above-mentioned nanodot aqueous solution 0.5mL prepared is taken, it is respectively 0,2nM, 10nM, 1 that 0.5mL concentration, which is added,
μM, 50 μM, 100 μM, 150 μM, 200 μM, 300 μM, 500 μM, the CuCl of 1mM, 2.5mM2Aqueous solution adds 2mL deionization
Aqueous solution.Above-mentioned solution is put into Fluorescence Spectrometer and tests its fluorescence intensity.Its fluorescence spectra shows, be added copper from
The fluorescence intensity of the increase of sub- concentration, gold nano point is gradually reduced, and copper ion concentration detection limit is calculated down to 0.7nM (figure
2a).And good linear relationship (Fig. 2 b) is presented between copper ion concentration and fluorescence intensity.
6 parts of above-mentioned each 0.5mL of nanodot aqueous solution prepared are taken, 2.5mL deionized water solution is separately added into
Its fluorescence intensity is tested, value is set as I0.Then the CuCl that 0.5mL concentration is 300 μM is separately added into every part of solution2、KCl、
NaCl、MgCl2、ZnCl2、FeCl3Aqueous solution, add 2mL deionized water.Above-mentioned solution is put into Fluorescence Spectrometer and is surveyed
Its fluorescence intensity is tried, value is set as I, draws (I0- I)/I0Column diagram (Fig. 3), it can be seen that Cu2+It can be by gold nano point
Fluorescence intensity is quenched 60% or more, and other impurities ion has little effect the fluorescence intensity of gold nano point.Show this
Detection method has very high selectivity for copper ion.
Embodiment 2
The NaOH aqueous solution that 0.5mL concentration is 1mol/L is separately added into 25mL deionized water, 10 μ L concentration are
The Bovine Serum Albumin in Aqueous Solution of 10mmol/L stirs 5min at 25 DEG C, and it is 50mmol/L's that 500 μ L concentration, which are then added,
HAuCl4 aqueous solution stirs 25min at 30 DEG C.Gold nanoparticle can be obtained.It is dense that gold nanoparticle aqueous solution is obtained at this time
Degree is 0.5mmol/L.
The gold nanoparticle that 10mL is prepared is taken, the NaOH aqueous solution that 50 μ L concentration are 1mol/L is added, adds
0.025g cysteine stirs 20h at 120 DEG C.Collect supernatant in eccentric fashion after reaction, and by supernatant weight
It is centrifuged 3 times again, finally obtaining supernatant is nanodot of the present invention.Under transmission electron microscope, gold nano spot size
For 2.2nm or so, uniform particle sizes, good dispersion.
The above-mentioned nanodot 0.5mL prepared is taken, it is respectively 0,2nM, 10nM, 1 μM, 50 μ that 0.5mL concentration, which is added,
M, 100 μM, 150 μM, 200 μM, 300 μM, 500 μM, the CuSO of 1mM, 2.5mM4Aqueous solution adds 2mL deionized water solution.
Above-mentioned solution is put into Fluorescence Spectrometer and tests its fluorescence intensity.Its fluorescence spectra is shown, with addition copper ion concentration
Increase, the fluorescence intensity of gold nano point is gradually reduced, and copper ion concentration detection limit is calculated down to 2nM.And copper ion is dense
Good linear relationship is presented between degree and fluorescence intensity.
6 parts of above-mentioned each 0.5mL of nanodot aqueous solution prepared are taken, 2.5mL deionized water solution is separately added into
Its fluorescence intensity is tested, value is set as I0.Then the CuCl that 0.5mL concentration is 300 μM is separately added into every part of solution2、KCl、
NaCl、MgCl2、ZnCl2、FeCl3Aqueous solution, add 2mL deionized water.Above-mentioned solution is put into Fluorescence Spectrometer and is surveyed
Its fluorescence intensity is tried, value is set as I, draws (I0- I)/I0Column diagram, it can be seen that Cu2+It can be strong by the fluorescence of gold nano point
Degree is quenched 80% or more, and other impurities ion has little effect the fluorescence intensity of gold nano point.Show the detection side
Method has very high selectivity for copper ion.
Embodiment 3
The NaOH aqueous solution that 0.5mL concentration is 1mol/L is separately added into 25mL deionized water, 10 μ L concentration are
The tetramethylol chloride aqueous solution of 10mmol/L, stirs 5min at 25 DEG C, and it is 50mmol/L's that 500 μ L concentration, which are then added,
HAuCl4 aqueous solution stirs 25min at 30 DEG C.Gold nanoparticle can be obtained.It is dense that gold nanoparticle aqueous solution is obtained at this time
Degree is 0.8mmol/L.
The gold nanoparticle that 10mL is prepared is taken, the NaOH aqueous solution that 50 μ L concentration are 1mol/L is added, adds
0.025g glutathione stirs 20h at 120 DEG C.Collect supernatant in eccentric fashion after reaction, and by supernatant weight
It is centrifuged 3 times again, finally obtaining supernatant is nanodot of the present invention.Under transmission electron microscope, gold nano spot size
For 2.5nm or so, uniform particle sizes, good dispersion (Fig. 1 b).
The above-mentioned nanodot 0.5mL prepared is taken, it is respectively 0,2nM, 10nM, 1 μM, 50 μ that 0.5mL concentration, which is added,
M, 100 μM, 150 μM, 200 μM, 300 μM, 500 μM, the CuSO of 1mM, 2.5mM4Aqueous solution adds 2mL deionized water solution.
Above-mentioned solution is put into Fluorescence Spectrometer and tests its fluorescence intensity.Its fluorescence spectra is shown, with addition copper ion concentration
Increase, the fluorescence intensity of gold nano point is gradually reduced, and copper ion concentration detection limit is calculated down to 1.2nM.And copper ion
Good linear relationship is presented between concentration and fluorescence intensity.
6 parts of above-mentioned each 0.5mL of nanodot aqueous solution prepared are taken, 2.5mL deionized water solution is separately added into
Its fluorescence intensity is tested, value is set as I0.Then the CuCl that 0.5mL concentration is 300 μM is separately added into every part of solution2、KCl、
NaCl、MgCl2、ZnCl2、FeCl3Aqueous solution, add 2mL deionized water.Above-mentioned solution is put into Fluorescence Spectrometer and is surveyed
Its fluorescence intensity is tried, value is set as I, draws (I0- I)/I0Column diagram, it can be seen that Cu2+It can be strong by the fluorescence of gold nano point
Degree is quenched 65% or more, and other impurities ion has little effect the fluorescence intensity of gold nano point.Show the detection side
Method has very high selectivity for copper ion.
Claims (5)
1. a kind of preparation method of blue-fluorescence gold nanodot solution, its step are as follows:
1) in 20~30mL deionized water, the NaOH aqueous solution that 0.25~1mL concentration is 0.01~1mol/L is added, is added dense
Degree is the polyfunctional group macromolecular aqueous solution of 0.001~1mol/L as stabilizer and reducing agent, wherein NaOH and polyfunctional group
The dosage molar ratio of macromolecular is 0.1~10:1；Then solution is stirred at 20~30 DEG C to 5~10min, then rapidly joined dense
Degree contains Au for 1~100mmol/L's3+Aqueous solution, polyfunctional group macromolecular and Au3+Dosage molar ratio be 10~1000:
1；Solution is finally stirred to 20~60min at 20~30 DEG C to get large-sized gold nanoparticle aqueous solution, Jenner's grain of rice is arrived
The concentration of son is 0.5~10mmol/L；
2) in the gold nanoparticle aqueous solution that concentration prepares for 0.5~10mmol/L, addition concentration is 1~100mmol/L
NaOH aqueous solution, add concentration be 0.01~1mol/L the biomolecule aqueous solution containing sulfydryl as stabilizer and etching
Agent；Wherein the dosage molar ratio of gold nanoparticle aqueous solution and the biomolecule containing sulfydryl be 0.01~0.1:1, NaOH and contain sulfydryl
The dosage molar ratio of biomolecule is 0.1~1:1；Solution is stirred to 20~30h at 80~150 DEG C again, after reaction with
The mode of centrifugation collects supernatant, and by supernatant repeated centrifugation 3~5 times, finally obtained supernatant is fluorescence gold nano
Point aqueous solution.
2. a kind of preparation method of blue-fluorescence gold nanodot solution as described in claim 1, it is characterised in that: polyfunctional group
Macromolecular aqueous solution is tetramethylol chloride aqueous solution or Bovine Serum Albumin in Aqueous Solution.
3. a kind of preparation method of blue-fluorescence gold nanodot solution as described in claim 1, it is characterised in that: containing sulfydryl
Biomolecule aqueous solution is aqueous cystein solution or glutathione aqueous solution.
4. a kind of blue-fluorescence gold nanodot solution, it is characterised in that: be the method as described in claims 1 to 3 Ren Xiangyi
It is prepared.
5. blue-fluorescence gold nanodot solution as claimed in claim 4 is in the application of bivalent cupric ion context of detection.
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