CN109211862A - A kind of preparation method and applications of red fluorescence copper nanocluster probe - Google Patents
A kind of preparation method and applications of red fluorescence copper nanocluster probe Download PDFInfo
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- CN109211862A CN109211862A CN201811235660.2A CN201811235660A CN109211862A CN 109211862 A CN109211862 A CN 109211862A CN 201811235660 A CN201811235660 A CN 201811235660A CN 109211862 A CN109211862 A CN 109211862A
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
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Abstract
The invention discloses a kind of preparation method and applications of red fluorescence copper nanocluster probe, the present invention is to be prepared using dopamine as protective agent and reducing agent by " one kettle way "；Compound concentration is the dopamine of 10-20 mmol/L first, is stirred continuously down, and the copper nitrate solution that concentration is 15-40 mmol/L is added into dopamine solution, and continuing stirring mixes well the two, and the mass ratio of the material of copper nitrate and dopamine is 3 ~ 16:4；By gained mixed solution heating stirring, stand at room temperature；Then by centrifugation, red fluorescence copper nanocluster probe solution is finally obtained.The method of the present invention is made uniform in size, stability is good, red fluorescence copper nanocluster；Red fluorescence copper nanocluster probe good water solubility obtained, stability are strong, can be applied to the detection of human serum albumins (HSA).
Description
Technical field
The present invention relates to a kind of preparation method and applications of red fluorescence copper nanocluster probe, belong to fluorescence nano material
Expect preparation field.
Background technique
Metal nanometer cluster is that one kind is made of tens atoms, extra small nanoparticle of the metallic core size less than 2 nm
Son.As novel fluorescence nano-probe, metal nanometer cluster have unique electricity, physics and optical property, in recent years by
Intensive report is for detecting plurality of target object.One obvious characteristic of metal nanometre cluster is its strong luminescence generated by light, and has
Size is small, nontoxic, good water solubility, Stocks are displaced the big and anti-light stronger feature of bleaching power, thus it is wide to cause researcher
General interest.For gold and silver, the cheaper of copper.Therefore, copper nanocluster is increasingly becoming metal nano material
In important component, and be widely used in the research fields such as chemical analysis, bio-sensing, bio-imaging, ion detection.
Currently, the copper nanocluster of most of synthesis emits blue light under ultraviolet excitation.In analysis detection and biology
Aspect, red fluorescence copper nanocluster more attractive thus can be to avoid the interference of some organism autofluorescences.
Biological micromolecule such as glutathione, cysteine and lauryl mercaptan etc. are used as synthesizing the excellent steady of copper nanocluster
Determine agent.Document (Cation-driven luminescent self-assembled dots of copper
nanoclusters with aggregation-induced emission for b-galactosidase activity
monitoring, Y.Y. Huang, H. Feng, W. D. Liu, S.S. Zhang, C. Tang, J.R. Chen,
Z.S. Qian, J. Mater. Chem. B 2017,5,5120-5127), with a kind of common biological micromolecule-paddy
The sweet peptide of Guang synthesizes copper nano-cluster, can carry out the detection to galactosidase.But this method synthesis process is a bit cumbersome.Document
(Copper nanoclusters as an on-off-on fluorescent probe for ascorbic acid, H.
B. Rao, H.W. Ge, Z.W. Lu, W. Liu, Z.Q. Chen, Z.Y. Zhang, X.X. Wang, P. Zou,
Y.Y. Wang, H. He, X.Y. Zeng, Microchim Acta .2016,183,1651-1657) synthesis copper receive
Rice cluster is used for the detection of ascorbic acid, and is successfully applied to detection such as cucumber, Xi Lanhua, the hardship of the rate of recovery in practical biological sample
Melon, however, synthesis process is a bit cumbersome, and there are certain interference for biological detection for the copper nanocluster of hair blue-fluorescence
Property.
Summary of the invention
The present invention is intended to provide a kind of preparation method and applications of red fluorescence copper nanocluster probe, preparation method letter
Single, one-step synthesis, reaction condition is mild, and gained red fluorescence copper nanocluster probe can be to avoid the dry of organism autofluorescence
It disturbs, the detection of HSA can be used for.
Since the surface area of metal nanometre cluster is high, the valence link height unsaturation of outer layer atom keeps its surface free energy very high,
This results in trend of the nano-cluster with automatic agglomeration.So stable metal nanometre cluster is made, during the preparation process,
It is generally necessary to which protective agent is added to reduce surface free energy so that nanocluster uniform in size, that stability is good is made.The present invention with
The copper nano-cluster of hair red fluorescence is made as protective agent and reducing agent in dopamine, and this method is easy to operate, and cost is relatively low, former
Material is easy to get extensively, has good repeatability.
The present invention provides a kind of red fluorescence copper nanocluster probe, be using dopamine as protective agent and reducing agent,
It is prepared by " one kettle way ".
The present invention provides a kind of preparation methods of red fluorescence copper nanocluster probe, comprising the following steps:
(1) compound concentration is the dopamine of 10-20 mmol/L, is stirred continuously down, and concentration is added into dopamine solution and is
The copper nitrate solution of 15-40 mmol/L, continuing stirring mixes well the two, and the mass ratio of the material of copper nitrate and dopamine is 3
~16:4；
(2) mixed solution for obtaining step (1), takes 2-4mL, and heating stirring 6-7 h, stands at room temperature at 55-70 DEG C
12h；
(3) mixed solution for obtaining step (2) finally obtains red fluorescence copper nanocluster probe solution by centrifugation.
In above-mentioned preparation method, the concentration of the dopamine solution in step (1) is 10-20 mmol/L；Dopamine solution
Volume ratio with copper nitrate solution is 1:1；
Temperature is preferably 65 DEG C in step (2), and mixing time is preferably 6h.
Centrifuge speed is 10000-13000 r/min in step (3)；Centrifugation time is 10-15 min.
The present invention provides using red fluorescence copper nanocluster probe made from above-mentioned preparation method.
The present invention provides above-mentioned red fluorescence copper nanocluster probe answering in human serum albumins (HSA) detection
With.
Specific application are as follows: red fluorescence copper nanocluster probe solution is added in secondary distilled water and is diluted, is taken dilute
100 μ L of red fluorescence copper nanocluster probe solution after releasing is added to 900 μ L containing the molten of different ions or other small molecules
In liquid, fixed excitation wavelength is 560 nm, fluorescence spectrum detection is carried out in 0-10 min at room temperature, according to 624 nm's or so
Fluorescence peak intensity, is detected.
Beneficial effects of the present invention:
(1) environmentally protective using dopamine as template, preparation method is simple, low in cost.
(2) red fluorescence copper nanocluster probe size is small made from, photostability is strong, toxic side effect is small, water-soluble
Good, fluorescence intensity is high, has broad application prospects in fields such as bio-imaging, biomarkers.
(3) fluorescence copper nanocluster probe made from has good red luminous performance, for it is actually detected when can be with
Avoid the interference of some organism autofluorescences.
Detailed description of the invention
Fig. 1 is the mechanism of action schematic diagram of the red fluorescence copper nanocluster probe of present invention preparation and application；
Fig. 2 is that the embodiment of the present invention 1 prepares the ultraviolet figure of fluorescence-of fluorescence copper nanocluster probe solution, in figure a be it is ultraviolet-can
See abosrption spectrogram, b is fluorescence spectra；
When Fig. 3 is that different ions and other small molecules are added in red fluorescence copper nanocluster probe solution in the embodiment of the present invention 3
The variation diagram of fluorescence peak intensity；
In Fig. 4 embodiment of the present invention 2 red fluorescence copper nanocluster probe solution with ionic strength (concentration of sodium chloride) change
Change the variation diagram of its fluorescence peak intensity；
The fluorescence intensity of red fluorescence copper nanocluster probe solution changes with time figure in Fig. 5 embodiment of the present invention 4；
In Fig. 6 embodiment of the present invention 5 red fluorescence copper nanocluster probe solution with dopamine concentration increased fluorescence intensity
Variation diagram；
The dopamine of the fluorescence intensity of red fluorescence copper nanocluster probe solution and various concentration in Fig. 7 embodiment of the present invention 5
The linear relationship chart of solution.
Specific embodiment
The present invention is further illustrated below by embodiment, but is not limited to following embodiment.
The present invention is to prepare red fluorescence copper nanocluster probe by " one kettle way " using dopamine as template and reducing agent
Solution, and the detection for dopamine in solution.Below by example combination attached drawing, the invention will be further described.
Embodiment 1
Using dopamine as the preparation of the red fluorescence copper nanocluster probe of template:
(1) dopamine for preparing 20 mmol/L, is stirred continuously down, and 25 mmol/L copper nitrate solutions are added into dopamine solution,
Continuing stirring mixes well the two, and the volume ratio of dopamine solution and copper nitrate solution is 1:1；The object of copper nitrate and dopamine
The amount ratio of matter is 5:4；
(2) mixed solution for obtaining step (1), volume are 2 mL heating stirring 6h at 65 DEG C, at room temperature, are stood
12h；
(3) mixed solution for obtaining step (2) finally obtains red fluorescence copper nanocluster probe solution by centrifugation.
The mechanism of action schematic diagram of the red fluorescence copper nanocluster probe of preparation is shown in Fig. 1.
The fluorescence copper nanocluster probe solution of preparation is dark-brown in daylight light irradiation hypostome color, ultraviolet in 365 nm
It is red under light irradiation.
In addition, the ultraviolet figure of fluorescence-of the fluorescence copper nanocluster probe solution of preparation is shown in Fig. 2, show the fluorescence copper of preparation
For nanocluster probe under the conditions of fixed excitation wavelength is 560 nm, emission peak positions are in 624 nm or so.
Embodiment 2
The shadow of the fluorescence peak intensity for the fluorescence copper nanocluster probe solution that zwitterion and other small molecules prepare embodiment 1
Ring experiment:
With secondary distilled water and NaNO3、KNO3、Mg(NO3)2、Ca(NO3)2、Hg(NO3)2、Zn(NO3)2、Cu(NO3)2、Al
(NO3)2、Co(NO3)2、Cr(NO3)2、KCl、KBr、KI、KAc、K2SO4, GSH, Cys, Hcy, AA be configured to concentration respectively and be
0.1mol·L-1Solution.Red fluorescence copper nanocluster probe solution prepared by embodiment 1 dilutes 10 times, after taking dilution
100 μ L of red fluorescence copper nanocluster probe solution is added to the above-mentioned solution containing different ions or other small molecules of 900 μ L
In, fixed excitation wavelength is 560 nm, fluorescence spectrum detection is carried out at room temperature, according to the fluorescence peak intensity of 624 nm or so,
Detect the influence (Fig. 3) of different ions or small molecule to the fluorescence peak intensity of red fluorescence copper nanocluster probe solution.
Fig. 4 is shown in the influence of the fluorescence peak intensity of ion pair red fluorescence copper nanocluster probe solution: exciting in 560 nm
Under, from fluorescence intensity F and red fluorescence copper nanoclusters containing ion or the red fluorescence copper nanocluster probe solution of small molecule
The fluorescence peak intensity F of aggregate probe solution0Ratio obtain: dopamine variation is maximum, other ions or small molecule variation are relatively
It is small, illustrate that red fluorescence copper nanocluster probe solution prepared by the present invention is able to detect dopamine.
Embodiment 3
The influence of the fluorescence peak intensity for the red fluorescence copper nanocluster probe solution that ionic strength prepares embodiment 1 is tested:
Red fluorescence copper nanocluster probe solution prepared by 100 μ L embodiments 1 is added in 900 μ L secondary distilled waters,
Fixed excitation wavelength is 560 nm, the sodium chloride solution (0.04-0.22 mol/L) of various concentration is added, according to 624 nm or so
Fluorescence peak intensity, detect influence of the ionic strength to the fluorescence peak intensity of red fluorescence copper nanocluster probe solution.
Fig. 3 is shown in influence of the ionic strength to the fluorescence peak intensity of fluorescence copper nanocluster probe solution: exciting in 560nm
Under, red fluorescence copper nanocluster probe solution is glimmering in sodium chloride solution (0.04-0.22 mol/L) range of various concentration
Photopeak intensity is basically unchanged, and illustrates that the anti-ion interference of red fluorescence copper nanocluster probe solution prepared by the present invention is strong.
Embodiment 4
Time red fluorescence copper nanocluster probe solution prepared by embodiment 1 is added the influence experiment after HSA:
After HSA is added in red fluorescence copper nanocluster probe solution prepared by 100 μ L embodiments 1, it is added to 900 μ L and goes
In ionized water, fixed excitation wavelength is 560 nm, carries out fluorescence spectrum detection in 0-10 min at room temperature, according to the left side 624 nm
Right fluorescence peak intensity, influence of the detection time to the fluorescence peak intensity of fluorescence copper nanocluster probe solution.Time is to fluorescence
Fig. 5 is shown in the influence of the fluorescence intensity of copper nanocluster probe solution: in 10 min, the fluorescence of fluorescence copper nanocluster probe is strong
Degree is held essentially constant.
Embodiment 5
The experiment that red fluorescence copper nanocluster probe solution prepared by embodiment 1 detects HSA:
Red fluorescence copper nanocluster probe solution prepared by embodiment 1 dilutes 10 times, the fluorescence copper nanoclusters after taking dilution
100 μ L of aggregate probe solution is added in 900 solution of the μ L containing HSA, and fixed excitation wavelength is 560 nm, is carried out at room temperature glimmering
Light spectral detection detects HSA to the fluorescence of red fluorescence copper nanocluster probe solution according to the fluorescence intensity of 624 nm or so
The influence of intensity.
Fig. 6 is shown in influence of the HSA to the fluorescence intensity of red fluorescence copper nanocluster probe solution: under 560 nm excitation,
Fluorescence copper nanocluster probe solution is after the HSA that various concentration is added, and fluorescence intensity is gradually reduced, and last fluorescence peak becomes substantially
In smooth；Wherein 0-200 μ g/mL is 0,1,5,10,15,20,40,60,80,100,120,140 respectively,
The fluorescence spectra that the HSA of 160,180,200 μ g/mL influences fluorescence copper nanocluster probe solution fluorescence intensity, explanation
Red fluorescence copper nanocluster probe solution prepared by the present invention can be realized the detection to HSA.
In addition, changing for the fluorescence intensity of red fluorescence copper nanocluster probe solution prepared by the present invention is dense with HSA
Spend it is in a linear relationship, as shown in fig. 7, the fluorescence intensity of copper nanocluster and the concentration of HSA are linear at two sections.(a) institute in Fig. 7
Show, the linear equation of HSA is F0/F=1.102+0.007C (R2=0.991)；In Fig. 7 shown in (b), the linear equation of HSA is
F0/F=-7.910+0.095C (R2=0.970)。
Claims (8)
1. a kind of preparation method of red fluorescence copper nanocluster probe, it is characterised in that: be using dopamine as protective agent and
Reducing agent is prepared by one kettle way.
2. the preparation method of red fluorescence copper nanocluster probe according to claim 1, it is characterised in that including following
Step:
(1) compound concentration is the dopamine of 10-20 mmol/L, is stirred continuously down, and concentration is added into dopamine solution and is
The copper nitrate solution of 15-40 mmol/L, continuing stirring mixes well the two；
(2) mixed solution for obtaining step (1), taking volume is that 2-4 mL stirs 6-7 h at 55-70 DEG C, is stood at room temperature
12 h；
(3) mixed solution for obtaining step (2) finally obtains red fluorescence copper nanocluster probe solution by centrifugation.
3. the preparation method of red fluorescence copper nanocluster probe according to claim 2, it is characterised in that: step (1)
In, the volume ratio of dopamine solution and copper nitrate solution is 1:1.
4. the preparation method of red fluorescence copper nanocluster probe according to claim 2, it is characterised in that: step (2)
Middle temperature is 65 DEG C.
5. the preparation method of red fluorescence copper nanocluster probe according to claim 2, it is characterised in that: step (3)
Middle centrifuge speed is 10000-13000 r/min；Centrifugation time is 10-15 min.
6. red fluorescence copper nanocluster probe made from a kind of described in any item preparation methods of claim 1 ~ 5.
7. a kind of application of red fluorescence copper nanocluster probe as claimed in claim 6 in human serum albumins detection.
8. application according to claim 7, it is characterised in that: red fluorescence copper nanocluster probe solution is added to two
Diluted in secondary distilled water, the 100 μ L of red fluorescence copper nanocluster probe solution after taking dilution be added to 900 μ L containing it is different from
In son or the solution of other small molecules, fixed excitation wavelength is 560 nm, carries out fluorescence spectrum inspection in 0-10 min at room temperature
It surveys, according to the fluorescence peak intensity of 624 nm, is detected.
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Cited By (3)
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| CN110724156A (en) * | 2019-10-22 | 2020-01-24 | 安徽大学 | Method for enhancing fluorescence intensity of copper nanocluster |
| CN111548792A (en) * | 2020-04-25 | 2020-08-18 | 山西大学 | Fluorescent copper nanocluster and preparation method and application thereof |
| CN112916863A (en) * | 2021-01-19 | 2021-06-08 | 山西大学 | Water-soluble luminescent silver nanocluster and preparation method and application thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110724156A (en) * | 2019-10-22 | 2020-01-24 | 安徽大学 | Method for enhancing fluorescence intensity of copper nanocluster |
| CN111548792A (en) * | 2020-04-25 | 2020-08-18 | 山西大学 | Fluorescent copper nanocluster and preparation method and application thereof |
| CN111548792B (en) * | 2020-04-25 | 2022-09-23 | 山西大学 | Fluorescent copper nanocluster and preparation method and application thereof |
| CN112916863A (en) * | 2021-01-19 | 2021-06-08 | 山西大学 | Water-soluble luminescent silver nanocluster and preparation method and application thereof |
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