CN110862820A - Preparation method and application of cysteine-gold nanocluster - Google Patents
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Abstract
The invention discloses a preparation method and application of cysteine-gold nanoclusters, and belongs to the technical field of fluorescence analysis. Adding a cysteine solution into ultrapure water under the stirring condition to obtain a solution A; adding chloroauric acid into the solution A under the condition of stirring, adding a sodium hydroxide solution, and uniformly mixing to obtain a solution B; and (3) reacting the solution B for 1-6 h at the temperature of 50-90 ℃ to obtain the weak-fluorescence Cys-Au nano-cluster. The invention utilizes tetrachloroauric acid (HAuCl)4) Cysteine (Cys) and sodium hydroxide (NaOH) are used as raw materials, the gold nanocluster with cysteine as a coating agent is synthesized, the gold nanocluster is weak in fluorescence, but the fluorescence of the gold nanocluster can be rapidly and remarkably enhanced by adding silver ions, the fluorescence color is yellow green, the maximum excitation peak of the fluorescence is 378nm, and the maximum emission peak is 535 nm. Based on the phenomenon, the rapid, high-selectivity and visual fluorescence analysis and detection of silver ions can be realized.
Description
Technical Field
The invention relates to a preparation method and application of cysteine-gold nanoclusters, and belongs to the technical field of fluorescence analysis.
Background
The high-selectivity and rapid detection of metal ions has important significance in environmental and food analysis. The Au-S bond between gold and sulfur is active in chemical nature and forms a rich compound material. These materials include oligomers coordinated by gold thiolates and supramolecular assemblies of gold, such as gold nanoclusters with well defined atoms, gold nanoparticles of slightly larger size protected by thiolate monolayers. The gold thiolate compound combines strong luminescence, low toxicity, ultrafine size and good biocompatibility, so that the gold thiolate compound has wide application prospects in the fields of biosensors, environmental monitoring, cell imaging, disease detection, drug delivery and the like.
There are many reports in the literature on methods for synthesizing gold mercaptide compounds. The chemical reduction method is more common, that is, a specific coating agent or template molecule is selected to coat metal ions, and then a reducing agent is used to reduce the gold ions to a low valence state. Common coating agents containing thiol groups are reduced glutathione, cysteine, lipoic acid and some thiol compounds. For example, ZHENtao Luo et al synthesizes the high-luminescence gold-thiolate nanoclusters by a aggregation-induced emission mechanism and a glutathione one-pot method. For example, Wenli Hou et al utilize Ag+The modification enhances the fluorescence of the glutathione-gold nanocluster, and the glutathione-gold nanocluster is used as a fluorescence quenching nanoprobe for the ultra-sensitive detection of iodide.
However, the application of the gold mercaptide compound is directly influenced by the luminescence property of the gold mercaptide compound, and although the synthesis methods reported in many literatures display luminescence in a blue to near infrared region, most of the gold mercaptide compounds have low fluorescence quantum yield; the method for detecting the biological molecules is based on a fluorescence quenching mechanism of gold thiolate compounds for detection, and has low sensitivity and unobvious visual detection phenomenon when low-concentration detection is carried out; in the field of heavy metal analysis, an analysis method using gold nanoclusters or gold nanoclusters as fluorescent probes is mainly based on fluorescence quenching, and is poor in efficiency and long in reaction time.
Disclosure of Invention
Aiming at the defect of luminescent property of aurothiolate compounds in the prior art, the invention provides a preparation method and application of Cys-Au nano-cluster, and the invention utilizes tetrachloroauric acid (HAuCl)4) Cysteine (Cys) and sodium hydroxide (NaOH) are used as raw materials, the cysteine is synthesized to be used as a coating agent gold nanocluster, the gold nanocluster is weak in fluorescence, but the fluorescence of the nanocluster can be rapidly and remarkably enhanced by adding silver ions, the fluorescence color is yellow green, the maximum excitation peak of the fluorescence is 378nm, and the maximum emission peak is 535 nm. Based on the phenomenon, the rapid, high-selectivity and visual fluorescence analysis and detection of silver ions can be realized.
A preparation method of cysteine-gold nanoclusters comprises the following specific steps:
(1) adding the cysteine solution into ultrapure water under the stirring condition to obtain a solution A;
(2) adding chloroauric acid into the solution A obtained in the step (1) under the condition of stirring, adding a sodium hydroxide solution, and uniformly mixing to obtain a solution B;
(3) and (3) reacting the solution B for 1-6 h at the temperature of 50-90 ℃ to obtain the weak-fluorescence Cys-Au nano-cluster.
The concentration of cysteine in the solution A is 1.25-2.0 mM.
The molar ratio of the chloroauric acid to the cysteine in the cysteine solution in the step (2) is 1 (1.25-2.0).
The concentration of the sodium hydroxide in the solution B in the step (2) is 0.45-0.65 mM.
The application of the weak-fluorescence Cys-Au nanocluster in silver ion detection is realized by adding Ag+After the reaction, the fluorescence is obviously enhanced, and the quantitative analysis of the silver ions can be realized by detecting the fluorescence change before and after the nano-cluster.
The detection method comprises the following steps:
(1) adding the weak-fluorescence Cys-Au nanoclusters into a buffer solution to prepare a weak-fluorescence Cys-Au nanocluster buffer solution with a preset concentration, and adding standard Ag with different concentrations and the same volume+The solution is added with constant volume to obtain a preset series of Ag+Rapidly shaking and uniformly mixing buffer solution with gradient concentration, reacting for 5-30 min, measuring the change of fluorescence intensity, and drawing standard fluorescence intensity-Ag+A graph of concentration relationship;
(2) adding the solution to be detected into the weak fluorescence Cys-Au nanocluster buffer solution obtained in the step (1) and fixing the volume to obtain the Ag to be detected+And (3) rapidly shaking and uniformly mixing the buffer solution, reacting for 5-30 min, measuring the fluorescence intensity, and comparing the fluorescence intensity value with the standard fluorescence intensity-Ag in the step (1)+The Ag of the solution to be measured can be obtained by comparing the concentration relational graph+The concentration is used for realizing the quantitative analysis of silver ions; wherein the volume of the solution to be measured and the standard Ag in the step (1)+The volume of the solution was the same.
The buffer solution is Britton-Robinson wide buffer solution, and the pH value is 6-10.
The invention has the beneficial effects that:
the invention utilizes tetrachloroauric acid (HAuCl)4) Cysteine (Cys) and sodium hydroxide (NaOH) are used as raw materials, gold nanoclusters with cysteine as a coating agent are synthesized, the gold nanoclusters are weak in fluorescence, but the fluorescence of the nanoclusters can be rapidly and remarkably enhanced by adding silver ions, the fluorescence color is yellow green, the maximum excitation peak of the fluorescence is 378nm, and the maximum emission peak is 535 nm; the rapid, high-selectivity and visual fluorescent quantitative analysis and detection of silver ions can be realized.
Drawings
FIG. 1 shows example 1Ag+Enhancing the spectrogram of Cys-Au nanocluster fluorescence;
FIG. 2 shows Cys-Au nanoclusters + Ag of example 1+The subsequent fluorescence spectrogram;
FIG. 3 shows Cys-Au nanoclusters + Ag of example 1+The later ultraviolet absorption spectrogram;
FIG. 4 is a transmission electron micrograph of Cys-Au nanoclusters of example 1;
FIG. 5 shows Cys-Au nanoclusters of example 1+Ag+Transmission electron microscopy images of;
FIG. 6 is a graph of the selectivity enhanced fluorescence spectrum of Cys-Au nanoclusters of example 1;
FIG. 7 is a histogram of the selectively enhanced fluorescence spectra of Cys-Au nanoclusters of example 1;
FIG. 8 shows exemplary Cys-Au nanoclusters for different concentrations of Ag+Enhanced fluorescence spectrogram of (1);
FIG. 9 shows standard fluorescence intensity-Ag for Cys-Au nanoclusters of example 1+And (4) a concentration relation graph.
Detailed Description
The present invention will be described in further detail with reference to specific embodiments, but the scope of the present invention is not limited to the description.
Example 1: a preparation method of cysteine-gold nanoclusters comprises the following specific steps:
(1) adding the cysteine Cys solution into ultrapure water under the stirring condition to obtain a solution A; wherein the concentration of cysteine Cys in solution A is 1.5 mM;
(2) adding chloroauric acid into the solution A obtained in the step (1) under the condition of stirring, adding a sodium hydroxide solution, and uniformly mixing to obtain a solution B; wherein the molar ratio of the chloroauric acid to the cysteine in the cysteine solution is 1:1.5, and the concentration of the sodium hydroxide in the solution B is 0.60 mM;
(3) placing the solution B at the temperature of 90 ℃ for reacting for 3h to obtain the weak-fluorescence Cys-Au nano-cluster;
Cys-Au nanocluster enhanced fluorescence detection Ag+：
Putting 200 mu L of Cys-Au nano-cluster solution into a 1.5mL centrifuge tube, adding 200 mu L of BR (pH 9.0) buffer solution to prepare weak-fluorescence Cys-Au nano-cluster buffer solution, and adding Ag+The solution is added to a constant volume of 1mL to obtain Ag with a concentration of 30 MuM+Rapidly shaking and uniformly mixing buffer solution with concentration, and measuring the fluorescence change after reacting for 5 min;
example Ag+The spectrogram of enhanced Cys-Au nanocluster fluorescence is shown in FIG. 1, wherein the abscissa represents wavelength and the ordinate represents fluorescence intensity; as can be seen from FIG. 1, when Cys, NaOH, Ag+After the three components are reacted with each other,there is no fluorescence. When Cys, NaOH, HAuCl4After the three react, the fluorescence of the reaction product is very weak. Only when Cys, NaOH, HAuCl4After the three react, Ag is added+Reaction, finding that fluorescence can be obviously enhanced; example Ag+The excitation emission spectrogram of enhanced Cys-Au nanocluster fluorescence is shown in FIG. 2, wherein the abscissa represents wavelength and the ordinate represents relative fluorescence intensity; as can be seen from FIG. 2, the maximum fluorescence excitation wavelength is 378nm, and the maximum emission wavelength is 535 nm; ag+The ultraviolet-absorption spectrogram for enhancing Cys-Au nanocluster fluorescence is shown in FIG. 3, wherein the abscissa represents wavelength and the ordinate represents absorption intensity, and as can be seen from FIG. 3, no surface plasmon resonance absorption peak of Au NPs exists at about 520nm, which indicates that Cys-Au NCs are synthesized experimentally; the Transmission Electron Micrograph (TEM) of the Cys-Au nanocluster of this example is shown in FIG. 4, and the Cys-Au nanocluster + Ag+The Transmission Electron Microscopy (TEM) is shown in FIG. 5, and from FIGS. 4 to 5, Cys-Au nanoclusters and Cys-Au nanoclusters + Ag+The dispersibility of the (B) is better;
example Ag+The spectrogram of selectively enhanced Cys-Au nanocluster fluorescence is shown in FIG. 6, wherein the abscissa represents metal ions and the ordinate represents relative fluorescence intensity; the Cys-Au nanocluster selectivity enhanced fluorescence spectrum histogram is shown in FIG. 7, and it can be seen from FIGS. 6-7 that Ag+Selectively enhancing the fluorescence of Cys-Au nano-clusters;
standard fluorescence intensity-Ag+And (3) plotting a concentration relation:
putting 200 mu L of Cys-Au nano-cluster solution into a 1.5mL centrifuge tube, adding 200 mu L of BR (pH 9.0) buffer solution to prepare weak-fluorescence Cys-Au nano-cluster buffer solution, and adding standard Ag with different concentrations and the same volume+The solution is added to a constant volume of 1mL to obtain a series of Ag+Rapidly shaking and uniformly mixing buffer solution with gradient concentration, reacting for 5-30 min, measuring the change of fluorescence intensity, and drawing standard fluorescence intensity-Ag+A graph of concentration relationship; this example Cys-Au nanoclusters with different concentrations of Ag+The change in fluorescence intensity after the action is shown in FIG. 8, in which the abscissa represents Ag+Concentration, ordinate represents fluorescence intensity, Cys-Au nanocluster standard fluorescence intensity-Ag+The graph of the concentration relationship is shown in figure 9,as can be seen from FIGS. 8 to 9, the Ag content in the system is varied+The concentration is increased, and the fluorescence intensity of the Cys-Au nano cluster is gradually enhanced; Cys-Au nanocluster enhanced fluorescence paired Ag+And carrying out visual detection.
Example 2: a preparation method of cysteine-gold nanoclusters comprises the following specific steps:
(1) adding the cysteine Cys solution into ultrapure water under the stirring condition to obtain a solution A; wherein the concentration of cysteine Cys in solution A is 1.25 mM;
(2) adding chloroauric acid into the solution A obtained in the step (1) under the condition of stirring, adding a sodium hydroxide solution, and uniformly mixing to obtain a solution B; wherein the molar ratio of the chloroauric acid to the cysteine in the cysteine solution is 1:1.25, and the concentration of the sodium hydroxide in the solution B is 0.60 mM;
(3) placing the solution B at the temperature of 90 ℃ for reacting for 3h to obtain the weak-fluorescence Cys-Au nano-cluster;
Cys-Au nanocluster enhanced fluorescence detection Ag+：
Putting 200 mu L of Cys-Au nano-cluster solution into a 1.5mL centrifuge tube, adding 200 mu L of BR (pH 9.0) buffer solution to prepare weak-fluorescence Cys-Au nano-cluster buffer solution, and adding Ag+The solution is added to a constant volume of 1mL to obtain Ag with a concentration of 30 MuM+Rapidly shaking and uniformly mixing buffer solution with concentration, reacting for 5-30 min, and measuring the fluorescence change of the buffer solution;
example Ag+The fluorescence of the Cys-Au nano cluster is selectively enhanced, the fluorescence color is yellow green, the maximum excitation wavelength of the fluorescence is 378nm, and the maximum emission wavelength is 535 nm.
Example 3: a preparation method of cysteine-gold nanoclusters comprises the following specific steps:
(1) adding the cysteine Cys solution into ultrapure water under the stirring condition to obtain a solution A; wherein the concentration of cysteine Cys in solution A is 1.75 mM;
(2) adding chloroauric acid into the solution A obtained in the step (1) under the condition of stirring, adding a sodium hydroxide solution, and uniformly mixing to obtain a solution B; wherein the molar ratio of the chloroauric acid to the cysteine in the cysteine solution is 1:1.75, and the concentration of the sodium hydroxide in the solution B is 0.60 mM;
(3) placing the solution B at the temperature of 90 ℃ for reacting for 3h to obtain the weak-fluorescence Cys-Au nano-cluster;
Cys-Au nanocluster enhanced fluorescence detection Ag+：
Putting 200 mu L of Cys-Au nano-cluster solution into a 1.5mL centrifuge tube, adding 200 mu L of BR (pH 9.0) buffer solution to prepare weak-fluorescence Cys-Au nano-cluster buffer solution, and adding Ag+The solution is added to a constant volume of 1mL to obtain Ag with a concentration of 30 MuM+Rapidly shaking and uniformly mixing buffer solution with concentration, reacting for 5-30 min, and measuring the fluorescence change of the buffer solution;
example Ag+The fluorescence of the Cys-Au nano cluster is selectively enhanced, the fluorescence color is yellow green, the maximum excitation wavelength of the fluorescence is 378nm, and the maximum emission wavelength is 535 nm.
Example 4: a preparation method of cysteine-gold nanoclusters comprises the following specific steps:
(1) adding the cysteine Cys solution into ultrapure water under the stirring condition to obtain a solution A; wherein the concentration of cysteine Cys in solution A is 1.50 mM;
(2) adding chloroauric acid into the solution A obtained in the step (1) under the condition of stirring, adding a sodium hydroxide solution, and uniformly mixing to obtain a solution B; wherein the molar ratio of the chloroauric acid to the cysteine in the cysteine solution is 1:1.50, and the concentration of the sodium hydroxide in the solution B is 0.55 mM;
(3) placing the solution B at the temperature of 90 ℃ for reacting for 3h to obtain the weak-fluorescence Cys-Au nano-cluster;
Cys-Au nanocluster enhanced fluorescence detection Ag+：
Putting 200 mu L of Cys-Au nano-cluster solution into a 1.5mL centrifuge tube, adding 200 mu L of BR (pH 9.0) buffer solution to prepare weak-fluorescence Cys-Au nano-cluster buffer solution, and adding Ag+The solution is added to a constant volume of 1mL to obtain Ag with a concentration of 30 MuM+Rapidly shaking and uniformly mixing buffer solution with concentration, reacting for 5-30 min, and measuring the fluorescence change of the buffer solution;
example Ag+Selectively enhance the fluorescence of Cys-Au nano-clusters, and the fluorescence color isYellow-green, with a fluorescence maximum excitation wavelength of 378nm and a maximum emission wavelength of 535 nm.
Example 5: a preparation method of cysteine-gold nanoclusters comprises the following specific steps:
(1) adding the cysteine Cys solution into ultrapure water under the stirring condition to obtain a solution A; wherein the concentration of cysteine Cys in solution A is 1.50 mM;
(2) adding chloroauric acid into the solution A obtained in the step (1) under the condition of stirring, adding a sodium hydroxide solution, and uniformly mixing to obtain a solution B; wherein the molar ratio of the chloroauric acid to the cysteine in the cysteine solution is 1:1.50, and the concentration of the sodium hydroxide in the solution B is 0.65 mM;
(3) placing the solution B at the temperature of 90 ℃ for reacting for 3h to obtain the weak-fluorescence Cys-Au nano-cluster;
Cys-Au nanocluster enhanced fluorescence detection Ag+：
Putting 200 mu L of Cys-Au nano-cluster solution into a 1.5mL centrifuge tube, adding 200 mu L of BR (pH 9.0) buffer solution to prepare weak-fluorescence Cys-Au nano-cluster buffer solution, and adding Ag+The solution is added to a constant volume of 1mL to obtain Ag with a concentration of 30 MuM+Rapidly shaking and uniformly mixing buffer solution with concentration, reacting for 5-30 min, and measuring the fluorescence change of the buffer solution;
example Ag+The fluorescence of the Cys-Au nano cluster is selectively enhanced, the fluorescence color is yellow green, the maximum excitation wavelength of the fluorescence is 378nm, and the maximum emission wavelength is 535 nm.
Example 6: a preparation method of cysteine-gold nanoclusters comprises the following specific steps:
(1) adding the cysteine Cys solution into ultrapure water under the stirring condition to obtain a solution A; wherein the concentration of cysteine Cys in solution A is 1.50 mM;
(2) adding chloroauric acid into the solution A obtained in the step (1) under the condition of stirring, adding a sodium hydroxide solution, and uniformly mixing to obtain a solution B; wherein the molar ratio of the chloroauric acid to the cysteine in the cysteine solution is 1:1.50, and the concentration of the sodium hydroxide in the solution B is 0.60 mM;
(3) placing the solution B at the temperature of 90 ℃ to react for 1h to obtain the weak-fluorescence Cys-Au nano-cluster;
Cys-Au nanocluster enhanced fluorescence detection Ag+：
Putting 200 mu L of Cys-Au nano-cluster solution into a 1.5mL centrifuge tube, adding 200 mu L of BR (pH 9.0) buffer solution to prepare weak-fluorescence Cys-Au nano-cluster buffer solution, and adding Ag+The solution is added to a constant volume of 1mL to obtain Ag with a concentration of 30 MuM+Rapidly shaking and uniformly mixing buffer solution with concentration, reacting for 5-30 min, and measuring the fluorescence change of the buffer solution;
example Ag+The fluorescence of the Cys-Au nano cluster is selectively enhanced, the fluorescence color is yellow green, the maximum excitation wavelength of the fluorescence is 378nm, and the maximum emission wavelength is 535 nm.
Example 7: a preparation method of cysteine-gold nanoclusters comprises the following specific steps:
(1) adding the cysteine Cys solution into ultrapure water under the stirring condition to obtain a solution A; wherein the concentration of cysteine Cys in solution A is 2.00 mM;
(2) adding chloroauric acid into the solution A obtained in the step (1) under the condition of stirring, adding a sodium hydroxide solution, and uniformly mixing to obtain a solution B; wherein the molar ratio of the chloroauric acid to the cysteine in the cysteine solution is 1:2.0, and the concentration of the sodium hydroxide in the solution B is 0.60 mM;
(3) placing the solution B at the temperature of 90 ℃ for reacting for 3h to obtain the weak-fluorescence Cys-Au nano-cluster;
Cys-Au nanocluster enhanced fluorescence detection Ag+：
Putting 200 mu L of Cys-Au nano-cluster solution into a 1.5mL centrifuge tube, adding 200 mu L of BR (pH 9.0) buffer solution to prepare weak-fluorescence Cys-Au nano-cluster buffer solution, and adding Ag+The solution is added to a constant volume of 1mL to obtain Ag with a concentration of 30 MuM+Rapidly shaking and uniformly mixing buffer solution with concentration, reacting for 5-30 min, and measuring the fluorescence change of the buffer solution;
example Ag+The fluorescence of the Cys-Au nano cluster is selectively enhanced, the fluorescence color is yellow green, the maximum excitation wavelength of the fluorescence is 378nm, and the maximum emission wavelength is 535 nm.
Example 8: a preparation method of cysteine-gold nanoclusters comprises the following specific steps:
(1) adding the cysteine Cys solution into ultrapure water under the stirring condition to obtain a solution A; wherein the concentration of cysteine Cys in solution A is 1.50 mM;
(2) adding chloroauric acid into the solution A obtained in the step (1) under the condition of stirring, adding a sodium hydroxide solution, and uniformly mixing to obtain a solution B; wherein the molar ratio of the chloroauric acid to the cysteine in the cysteine solution is 1:1.50, and the concentration of the sodium hydroxide in the solution B is 0.60 mM;
(3) placing the solution B at the temperature of 80 ℃ for reacting for 6h to obtain the weak-fluorescence Cys-Au nano-cluster;
Cys-Au nanocluster enhanced fluorescence detection Ag+：
Putting 200 mu L of Cys-Au nano-cluster solution into a 1.5mL centrifuge tube, adding 200 mu L of BR (pH 9.0) buffer solution to prepare weak-fluorescence Cys-Au nano-cluster buffer solution, and adding Ag+The solution is added to a constant volume of 1mL to obtain Ag with a concentration of 30 MuM+Rapidly shaking and uniformly mixing buffer solution with concentration, reacting for 5-30 min, and measuring the fluorescence change of the buffer solution;
example Ag+The fluorescence of the Cys-Au nano cluster is selectively enhanced, the fluorescence color is yellow green, the maximum excitation wavelength of the fluorescence is 378nm, and the maximum emission wavelength is 535 nm.
Claims (5)
1. A preparation method of cysteine-gold nanoclusters is characterized by comprising the following specific steps:
(1) adding the cysteine solution into ultrapure water under the stirring condition to obtain a solution A;
(2) adding chloroauric acid into the solution A obtained in the step (1) under the condition of stirring, adding a sodium hydroxide solution, and uniformly mixing to obtain a solution B;
(3) and (3) reacting the solution B for 1-6 h at the temperature of 50-90 ℃ to obtain the weak-fluorescence Cys-Au nano-cluster.
2. The method of preparing cysteine-gold nanoclusters of claim 1, wherein: the concentration of cysteine in the solution A is 1.25-2.0 mM.
3. The method of preparing cysteine-gold nanoclusters of claim 1, wherein: the molar ratio of the chloroauric acid to the cysteine in the cysteine solution in the step (2) is 1 (1.25-2.0).
4. The method of preparing cysteine-gold nanoclusters of claim 1, wherein: the concentration of sodium hydroxide in the solution B is 0.45-0.65 mM.
5. The application of the Cys-Au nanocluster with weak fluorescence prepared by the preparation method of the cysteine-gold nanocluster as claimed in any one of claims 1 to 4 in silver ion detection.
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