CN103551587B - The preparation method of fluorescent nanometer silver cluster - Google Patents

The preparation method of fluorescent nanometer silver cluster Download PDF

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CN103551587B
CN103551587B CN201310515552.1A CN201310515552A CN103551587B CN 103551587 B CN103551587 B CN 103551587B CN 201310515552 A CN201310515552 A CN 201310515552A CN 103551587 B CN103551587 B CN 103551587B
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solution
silver
preparation
serum albumin
bovine serum
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CN103551587A (en
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郭志
陈桂秋
曾光明
晏铭
陈安伟
左亚男
黄真真
谭琼
刘灵芝
易斌
牛秋雅
张企华
黄健
杜坚坚
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Hunan University
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Abstract

The invention discloses a kind of preparation method of fluorescent nanometer silver cluster, the complex silver Na that the method generates with silver nitrate and sodium thiosulfate reaction 3[Ag (S 2o 3) 2] be Ag source, take bovine serum albumin(BSA) as reducing agent and stabilizing agent, by complex silver Na 3[Ag (S 2o 3) 2] add in bovine serum albumin solution, and regulate pH value to 10.46 ~ 14 of gained mixed solution, under normal temperature, lucifuge carries out oscillating reactions, obtains fluorescent nanometer silver cluster.The method is simple to operate, with low cost, reaction condition is gentle, favorable reproducibility and environmentally friendly.

Description

The preparation method of fluorescent nanometer silver cluster
Technical field
The invention belongs to materials chemistry and biology techniques field, be specifically related to a kind of preparation method of fluorescent nanometer silver cluster.
Background technology
The nano-Ag particles that Nano Silver (Nano Silver) cluster is less than Fermi's radius by particle diameter forms.The particle diameter of Nano Silver, mostly at 25 ran, has strong suppression and killing action to tens of kinds of pathogenic microorganisms such as Escherichia coli, gonococcus, chlamydia trachomatises, and can not produce drug resistance.Research about Nano Silver mainly concentrates on the antibiotic property of Nano Silver at present, its antibacterial feature has: broad-spectrum antiseptic, strong bactericidal, permeability are strong, reparative regeneration, antibacterial lasting, have no drug resistance, due to these features of Nano Silver, make its extensive application on modern medicine.
The energy level splitting that Nano Silver cluster produces because size is little, quantum size effect, make it have obvious fluorescent effect.In recent years, the relevant report about Nano Silver cluster is less, and preparation method is more complicated, fluorescence intensity and biocompatibility poor, domain size distribution heterogeneity, greatly reduces feasibility and validity that Nano Silver cluster applies in biological and medical science.
In view of nano metal material has slowly stepped into the field of life science, develop the preparation method of the high-quality Nano Silver cluster that a kind of technique is simple, Product size is even, particle dispersion is good, fluorescence intensity is high, reproducible, significant in the application of biological field to Nano Silver cluster.
Summary of the invention
The technical problem to be solved in the present invention overcomes the deficiencies in the prior art, there is provided a kind of simple to operate, with low cost, reaction condition is gentle, favorable reproducibility and the preparation method of environment amenable fluorescent nanometer silver cluster, the method prepare fluorescent nanometer silver cluster size uniformity, fluorescence intensity is high and fluorescent stability good.
For solving the problems of the technologies described above, the technical solution used in the present invention is a kind of preparation method of fluorescent nanometer silver cluster, the complex silver Na generated with silver nitrate and sodium thiosulfate reaction 3[Ag (S 2o 3) 2] be Ag source, take bovine serum albumin(BSA) as reducing agent and stabilizing agent, by complex silver Na 3[Ag (S 2o 3) 2] add in bovine serum albumin solution, and regulate pH value to 10.46 ~ 14 of gained mixed solution, under normal temperature, lucifuge carries out oscillating reactions, obtains fluorescent nanometer silver cluster.
Above-mentioned preparation method specifically comprises the following steps:
(1) under agitation, liquor argenti nitratis ophthalmicus is dropped in hypo solution, generate complex silver Na through reaction 3[Ag (S 2o 3) 2], after having reacted, obtain complex silver solution;
(2) under agitation, drop in bovine serum albumin solution by complex silver solution, then the pH value of gained mixed solution is adjusted to 10.46 ~ 14, under normal temperature, lucifuge carries out oscillating reactions, has reacted rear filtration, has obtained fluorescent nanometer silver cluster.
In above-mentioned preparation method, preferably, in described step (1), after obtaining complex silver solution, complex silver solution is continued to stir 1min ~ 3min.
In above-mentioned preparation method, preferably, described pH value is 11.25 ~ 13.75.
In above-mentioned preparation method, preferably, the ratio of the amount of substance (i.e. mol ratio) of described silver nitrate and described sodium thiosulfate is 1: 2.25 ~ 8.25.
In above-mentioned preparation method, preferably, in described mixed solution, the mass concentration of bovine serum albumin(BSA) is 0.25g/L ~ 1.77g/L.
In above-mentioned preparation method, preferably, the condition of described oscillating reactions is: reaction time 24h ~ 48h, rotating speed 145rpm ~ 175rpm.
In above-mentioned preparation method, preferably, described pH value is regulated by NaOH solution.
Compared with prior art, the invention has the advantages that:
Method of the present invention adopts nontoxic and the bovine serum albumin(BSA) of good biocompatibility is reducing agent and stabilizing agent prepares fluorescent nanometer silver cluster, overcomes and uses NaBH 4the bio-toxicity caused as reducing agent Deng chemical reagent and incompatibility, the biocompatibility of Nano Silver cluster prepared by the present invention has had large increase, preparation process of the present invention is carried out at normal temperatures, do not need special reaction condition, the size of fluorescence nano Argent grain can be controlled well by control ph, simple to operate, with low cost, there is good reappearance, be prepare low toxicity, geomery is homogeneous, fluorescence intensity is high, the green approach of the fluorescent nanometer silver cluster of fluorescent stability and good biocompatibility.
Accompanying drawing explanation
Fig. 1 is that in the embodiment of the present invention 1, the mass concentration of bovine serum albumin(BSA) in mixed solution is the transmission electron microscope photo of 0.88g/L, the fluorescent nanometer silver cluster prepared when pH value is 11.74.
Fig. 2 is that in the embodiment of the present invention 2, the mass concentration of bovine serum albumin(BSA) in mixed solution is the transmission electron microscope photo of 0.88g/L, the fluorescent nanometer silver cluster prepared when pH value is 12.08.
Fig. 3 is that in the embodiment of the present invention 2, the mass concentration of bovine serum albumin(BSA) in mixed solution is the grain size distribution of 0.88g/L, the fluorescent nanometer silver cluster prepared when pH value is 12.08.
Fig. 4 is the embodiment of the present invention 1, the mass concentration of bovine serum albumin(BSA) in mixed solution is 0.88g/L, the fluorescence emission spectrogram of the fluorescent nanometer silver cluster prepared when pH value is respectively 11.74,12.08,12.75, wherein λ in embodiment 2, embodiment 3 ex=462nm, CPS are fluorescence intensity unit, full name Counts Per Second.
Fig. 5 is the embodiment of the present invention 4, the mass concentration of bovine serum albumin(BSA) in mixed solution is respectively 0.44g/L, 0.88g/L, 1.32g/L in embodiment 5, embodiment 6, the fluorescence emission spectrogram of the fluorescent nanometer silver cluster prepared when pH value is 12.50, wherein λ ex=462nm.
Detailed description of the invention
Below in conjunction with Figure of description and concrete preferred embodiment, the invention will be further described, but protection domain not thereby limiting the invention.Normal temperature in the embodiment of the present invention is usually at 25 DEG C ~ 35 DEG C.
embodiment 1:
A preparation method for fluorescent nanometer silver cluster of the present invention, comprises the following steps:
(1) Na that 100ml concentration is 150mmol/L is got 2s 2o 383ml concentration, under the condition of vigorous stirring, is the AgNO of 74mmol/L by solution 3dropwise joins Na 2s 2o 3in solution, generate complex silver Na through reaction 3[Ag (S 2o 3) 2] (i.e. Ag source), after having reacted, obtain complex silver solution, and complex silver solution is continued to stir 1min;
(2) under agitation, above-mentioned complex silver solution 24ml is dropped to bovine serum albumin solution (the BSA solution that 10ml mass concentration is 10g/L, for reducing agent and stabilizing agent) in, then add a certain amount of sterilized water, make the final mass concentration of bovine serum albumin(BSA) in gained mixed solution be 0.88g/L; Regulate the pH value to 11.74 of mixed solution by the NaOH solution of 10mg/L, shake up and be placed in shaken cultivation case, under normal temperature, lucifuge carries out oscillating reactions 48h, rotating speed is 175rpm, after having reacted, adopts ordinary filter paper to filter product, collect filtrate, obtain fluorescent nanometer silver cluster.
As shown in Figure 1, be the transmission electron microscope photo of fluorescent nanometer silver cluster prepared by the present embodiment, from photo, gained Nano Silver cluster is uniformly dispersed, and particle size is consistent.
embodiment 2:
A preparation method for fluorescent nanometer silver cluster of the present invention, comprises the following steps:
(1) Na that 100ml concentration is 150mmol/L is got 2s 2o 383ml concentration, under the condition of vigorous stirring, is the AgNO of 74mmol/L by solution 3dropwise joins Na 2s 2o 3in solution, generate complex silver Na through reaction 3[Ag (S 2o 3) 2], after having reacted, obtain complex silver solution, and complex silver solution is continued to stir 1min;
(2) under the condition stirred, it is in the bovine serum albumin solution of 10g/L that above-mentioned complex silver solution 24ml is dropped to 10ml mass concentration, then add a certain amount of sterilized water, make the final mass concentration of bovine serum albumin(BSA) in gained mixed solution be 0.88g/L; Regulate the pH value to 12.08 of mixed solution by the NaOH solution of 10mg/L, shake up and be placed in shaken cultivation case, under normal temperature, lucifuge carries out oscillating reactions 48h, rotating speed is 175rpm, after having reacted, adopts ordinary filter paper to filter product, collect filtrate, obtain fluorescent nanometer silver cluster.
As shown in Figure 2, be the transmission electron microscope photo of fluorescent nanometer silver cluster prepared by the present embodiment, from photo, the particle size uniformity of gained fluorescent nanometer silver cluster, particle diameter is within the scope of 4 ~ 6nm.As shown in Figure 3, be the grain size distribution (being recorded by nanometer particle size potentiometric analyzer) of fluorescent nanometer silver cluster prepared by the present embodiment, as seen from the figure, the size distribution of fluorescence nano Argent grain is narrower, and average grain diameter is 10nm.Two kinds of Different Results cause due to apparatus measures principle difference, usually two kinds of methods are integrated and carry out granularmetric analysis.
embodiment 3:
A preparation method for fluorescent nanometer silver cluster of the present invention, comprises the following steps:
(1) Na that 100ml concentration is 150mmol/L is got 2s 2o 383ml concentration, when vigorous stirring, is the AgNO of 74mmol/L by solution 3dropwise joins Na 2s 2o 3in solution, generate complex silver Na through reaction 3[Ag (S 2o 3) 2], after having reacted, obtain complex silver solution, and complex silver solution is continued to stir 1min;
(2) under the condition stirred, it is in the bovine serum albumin solution of 10g/L that above-mentioned complex silver solution 24ml is dropped to 10ml mass concentration, then add a certain amount of sterilized water, make the final mass concentration of bovine serum albumin(BSA) in gained mixed solution be 0.88g/L; Regulate the pH value to 12.75 of mixed solution by the NaOH solution of 10mg/L, shake up and be placed in shaken cultivation case, under normal temperature, lucifuge carries out oscillating reactions 48h, rotating speed is 175rpm, after having reacted, adopts ordinary filter paper to filter product, collect filtrate, obtain fluorescent nanometer silver cluster.
Embodiment 1, embodiment 2, embodiment 3(and pH are respectively 11.74,12.08,12.75) the fluorescent nanometer silver cluster fluorescence emission spectrometer prepared tests, the result obtained as shown in Figure 4, as seen from the figure, Nano Silver cluster fluoresced green, with the rising of pH value, fluorescence intensity increases, and shows that the output of fluorescent nanometer silver cluster raises with pH value and increases to some extent.
embodiment 4:
A preparation method for fluorescent nanometer silver cluster of the present invention, comprises the following steps:
(1) Na that 100ml concentration is 150mmol/L is got 2s 2o 383ml concentration, under the condition of vigorous stirring, is the AgNO of 74mmol/L by solution 3dropwise joins Na 2s 2o 3in solution, generate complex silver Na through reaction 3[Ag (S 2o 3) 2], after having reacted, obtain complex silver solution, and complex silver solution is continued to stir 1min;
(2) under the condition stirred, above-mentioned complex silver solution 24ml being dropped to 5ml mass concentration is in the bovine serum albumin solution of 10g/L, then adds a certain amount of sterilized water, makes the final mass concentration of bovine serum albumin(BSA) in gained mixed solution be 0.44g/L; And the pH value to 12.50 of mixed solution is regulated by the NaOH solution of 10mg/L, shake up and be placed in shaken cultivation case, under normal temperature, lucifuge carries out oscillating reactions 48h, rotating speed is 175rpm, adopt ordinary filter paper to filter product, collect filtrate, obtain fluorescent nanometer silver cluster.
embodiment 5:
A preparation method for fluorescent nanometer silver cluster of the present invention, comprises the following steps:
(1) Na that 100ml concentration is 150mmol/L is got 2s 2o 383ml concentration, under the condition of vigorous stirring, is the AgNO of 74mmol/L by solution 3dropwise joins Na 2s 2o 3in solution, generate complex silver Na through reaction 3[Ag (S 2o 3) 2], after having reacted, obtain complex silver solution, and complex silver solution is continued to stir 1min;
(2) under the condition stirred, above-mentioned complex silver solution 24ml being dropped to 10ml mass concentration is in the bovine serum albumin solution of 10g/L, then adds a certain amount of sterilized water, makes the final mass concentration of bovine serum albumin(BSA) in gained mixed solution be 0.88g/L; Regulate the pH value to 12.50 of mixed solution by the NaOH solution of 10mg/L, shake up and be placed in shaken cultivation case, under normal temperature, lucifuge carries out oscillating reactions 48h, rotating speed is 175rpm, adopt ordinary filter paper to filter product, collect filtrate, obtain fluorescent nanometer silver cluster.
As shown in Figure 5, the maximum fluorescence intensity of gained Nano Silver cluster is about 300000cps, and fluorescence intensity is higher, and for characterizing its fluorescent stability, after placing three months to sample, survey its fluorescence intensity, its intensity does not change.
embodiment 6:
A preparation method for fluorescent nanometer silver cluster of the present invention, comprises the following steps:
(1) Na that 100ml concentration is 150mmol/L is got 2s 2o 383ml concentration, under the condition of vigorous stirring, is the AgNO of 74mmol/L by solution 3dropwise joins Na 2s 2o 3in solution, generate complex silver Na through reaction 3[Ag (S 2o 3) 2], after having reacted, obtain complex silver solution, and complex silver solution is continued to stir 1min;
(2) under the condition stirred, it is in the bovine serum albumin solution of 10g/L that above-mentioned complex silver solution 24ml is dropped to 15ml mass concentration, then a certain amount of sterilized water is added, the final mass concentration of bovine serum albumin(BSA) in gained mixed solution is made to be 1.32g/L, the pH value to 12.50 of mixed solution is regulated by the NaOH solution of 10mg/L, shake up and be placed in shaken cultivation case, under normal temperature, lucifuge carries out oscillating reactions 48h, rotating speed is 175rpm, ordinary filter paper is adopted to filter product, collect filtrate, obtain fluorescent nanometer silver cluster.
Embodiment 4, embodiment 5, the embodiment 6(bovine serum albumin(BSA) final mass concentration in mixed solution is respectively 0.44g/L, 0.88g/L, 1.32g/L) the fluorescent nanometer silver cluster fluorescence emission spectrometer prepared tests, the result obtained as shown in Figure 5, as seen from the figure, prepared fluorescent nanometer silver cluster has most hyperfluorescence emission peak at 550nm place, fluoresced green, along with the increase of bovine serum albumin(BSA) addition, the fluorescence intensity of fluorescent nanometer silver cluster increases, and shows that the output of fluorescent nanometer silver cluster increases to some extent.
embodiment 7:
A preparation method for fluorescent nanometer silver cluster of the present invention, comprises the following steps:
(1) Na that 100ml concentration is 150mmol/L is got 2s 2o 325ml concentration, under the condition of vigorous stirring, is the AgNO of 74mmol/L by solution 3dropwise joins Na 2s 2o 3in solution, now generate complex silver Na through reaction 3[Ag (S 2o 3) 2], after having reacted, obtain complex silver solution, and complex silver solution is continued to stir 1min;
(2) under the condition stirred, it is in the bovine serum albumin solution of 10g/L that above-mentioned complex silver solution 24ml is dropped to 10ml mass concentration, then add a certain amount of sterilized water, make the final mass concentration of bovine serum albumin(BSA) in gained mixed solution be 0.88g/L; And the pH value to 12.50 of mixed solution is regulated by the NaOH solution of 10mg/L, shake up and be placed in shaken cultivation case, under normal temperature, lucifuge carries out oscillating reactions 48h, rotating speed is 175rpm, adopt ordinary filter paper to filter product, collect filtrate, obtain fluorescent nanometer silver cluster.
The above is only the preferred embodiment of the present invention, and protection scope of the present invention is also not only confined to above-described embodiment.All technical schemes belonged under thinking of the present invention all belong to protection scope of the present invention.It is noted that for those skilled in the art, improvements and modifications under the premise without departing from the principles of the invention, these improvements and modifications also should be considered as protection scope of the present invention.

Claims (5)

1. a preparation method for fluorescent nanometer silver cluster, is characterized in that, the complex silver Na generated with silver nitrate and sodium thiosulfate reaction 3[Ag (S 2o 3) 2] be Ag source, take bovine serum albumin(BSA) as reducing agent and stabilizing agent, by complex silver Na 3[Ag (S 2o 3) 2] add in bovine serum albumin solution, and regulate pH value to 10.46 ~ 14 of gained mixed solution, under normal temperature, lucifuge carries out oscillating reactions, obtains fluorescent nanometer silver cluster;
Specifically comprise the following steps:
(1) under agitation, liquor argenti nitratis ophthalmicus is dropped in hypo solution, generate complex silver Na through reaction 3[Ag (S 2o 3) 2], after having reacted, obtain complex silver solution;
(2) under agitation, drop in bovine serum albumin solution by complex silver solution, then the pH value of gained mixed solution is adjusted to 10.46 ~ 14, under normal temperature, lucifuge carries out oscillating reactions, has reacted rear filtration, has obtained fluorescent nanometer silver cluster;
Described silver nitrate is 1: 2.25 ~ 8.25 with the ratio of the amount of substance of described sodium thiosulfate;
The condition of described oscillating reactions is: reaction time 24h ~ 48h, rotating speed 145rpm ~ 175rpm.
2. preparation method according to claim 1, is characterized in that, in described step (1), after obtaining complex silver solution, is continued to stir 1min ~ 3min by complex silver solution.
3. preparation method according to claim 1, is characterized in that, described pH value is 11.25 ~ 13.75.
4. preparation method according to claim 1, is characterized in that, in described mixed solution, the mass concentration of bovine serum albumin(BSA) is 0.25g/L ~ 1.77g/L.
5. preparation method according to claim 1, is characterized in that, described pH value is regulated by NaOH solution.
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CN104722776B (en) * 2015-04-09 2018-03-06 中南大学 A kind of silver receives the synthesis technique of cluster
CN106424753B (en) * 2016-10-14 2018-12-07 泉州师范学院 A kind of MnO2The preparation and its application of-Ag nanocomposite
CN112675198B (en) * 2019-10-18 2023-02-28 中国疾病预防控制中心病毒病预防控制所 Preparation method of novel silver nanocluster-ferritin bionic drug, drug and application
CN111588735A (en) * 2020-05-29 2020-08-28 西南大学 Application of nano silver in preparation of silkworm microparticle medicament for inhibiting main pathogen of silkworms
CN112897472B (en) * 2021-02-03 2022-09-06 晋大纳米科技（厦门）有限公司 Preparation method of high-temperature-resistant and illumination-resistant silver complexing solution, silver complexing solution and antibacterial product

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CN101559972A (en) * 2009-05-07 2009-10-21 上海交通大学 Preparation method of silver sulfide nano particle
CN102059344A (en) * 2010-11-11 2011-05-18 厦门大学 Method for synthesizing fluorescent silver nano-particles under microwave by polymethylacrylic acid
CN102590166A (en) * 2012-02-10 2012-07-18 中国科学院长春应用化学研究所 Test method for trypsin

Patent Citations (3)

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Publication number Priority date Publication date Assignee Title
CN101559972A (en) * 2009-05-07 2009-10-21 上海交通大学 Preparation method of silver sulfide nano particle
CN102059344A (en) * 2010-11-11 2011-05-18 厦门大学 Method for synthesizing fluorescent silver nano-particles under microwave by polymethylacrylic acid
CN102590166A (en) * 2012-02-10 2012-07-18 中国科学院长春应用化学研究所 Test method for trypsin

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