CN103264165B - A kind of method of synthesizing silver nanoclusters taking single stranded DNA as template - Google Patents

A kind of method of synthesizing silver nanoclusters taking single stranded DNA as template Download PDF

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CN103264165B
CN103264165B CN201310141932.3A CN201310141932A CN103264165B CN 103264165 B CN103264165 B CN 103264165B CN 201310141932 A CN201310141932 A CN 201310141932A CN 103264165 B CN103264165 B CN 103264165B
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single stranded
stranded dna
silver nanoclusters
template
method
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CN201310141932.3A
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CN103264165A (en
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冯九菊
周丹玲
奚焕祥
蔡琳熠
王爱军
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浙江师范大学
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Abstract

The invention provides a kind of method of synthesizing silver nanoclusters taking single stranded DNA as template, belong to technical field of nano material. It has solved the existing Nano Silver for detection of mercury ion and has produced complexity, the problem of high cost. Method of this synthetic silver nanoclusters taking single stranded DNA as template be the single stranded DNA that possesses cytimidine and guanine be template, with Ag+After solution mixes, add after reducing agent reduces and make the silver nanoclusters with fluorescence. In the present invention, we have set up a kind of simple, low cost, green taking commercially available polyinosinic acid (PI) and poly (PC) as template, method is synthesized silver nanoclusters reliably. Fluorescence utilizes, and as polyinosinic acid and the synthetic fluorescence silver nanoparticle crystalline substance of cytidine monophosphate, does not need special design and pretreatment. This novel silver nanoclusters has good selective and sensitivity, can be applicable to detect Hg2+, be feasible and have developing prospect for monitoring pollutant in water.

Description

A kind of method of synthesizing silver nanoclusters taking single stranded DNA as template

Technical field

The invention belongs to technical field of nano material, relate to a kind of silver nanoclusters, particularly a kind of method of synthesizing silver nanoclusters taking single stranded DNA as template.

Background technology

Environmental problem is people's new century to pay close attention to the most one of topic, and water pollution is the major issue that solution is badly in need of in environmental pollution. In water, polluting is exactly at most the harm of heavy metal, and heavy metal Hg exists with inorganic mercury or organomercurial form at nature, by the accumulation of biological chain, and the enrichment in vivo of mercury metal. Mercury and mercuric compounds to the infringement of human body with to enter mercury amount in body relevant. Mercury mainly involves central nervous system, digestive system and kidney to the harm of human body, in addition respiratory system, skin, blood and eyes is also had a certain impact. US and European is extremely low to the content requirement of Mercury in Drinking Water, must reach nanomole rank.

In the past few decades, people have set up many for quantitative assay Hg2+Method, comprise electrochemical method, atomic emission spectrometry, cold atomic absorption spectrometry, colorimetric method, inductively coupled plasma mass spectrometry and atomic fluorescence spectrometry. These methods require great effort relatively, time-consuming, somewhat expensive and need the sample treatment of multistep.

Molecular fluorescence method is with its high sensitivity and high selectivity and without advantages such as the instruments of any costliness and receive people's concern. But its needs well-designed, the synthetic and purifying of special mechanism, causes cost high, therefore cannot be used for conventional application.

Summary of the invention

The object of the invention is for detecting Hg in prior art2+With the problem of inconvenient, the high cost of silver nanoclusters preparation, and proposed a kind of taking single stranded DNA as template the method for synthetic silver nanoclusters, it has simply, cost is low, green reliable advantage.

Object of the present invention can realize by following technical proposal: a kind of taking single stranded DNA as template the method for synthetic silver nanoclusters, it is characterized in that, the method be the single stranded DNA that possesses cytimidine and guanine be template, with Ag+After solution mixes, add after reducing agent reduces and make the silver nanoclusters with fluorescence. With respect to bases adenine and thymidine, cytimidine and guanine and Ag+There is stronger affinity.

In a kind of above-mentioned method of synthesizing silver nanoclusters taking single stranded DNA as template, the concrete steps of described method comprise:

A, get a certain amount of single stranded DNA and Ag respectively+Solution, joins in cushioning liquid and mixes, and after mixing, waits for that certain hour enters step b again;

B, in the mixed solution in step a, add reducing agent, reduction obtains water soluble fluorescence silver nanoclusters product.

In step a, the reason that the rear wait of mixing certain hour enters step b is again: being in order to mix fully on the one hand, is to make it fully cooling under condition of ice bath on the other hand, thereby at NaBH4Reduction Ag+Reaction speed slow down, prevent generate too oarse-grained Ag.

In a kind of above-mentioned method of synthesizing silver nanoclusters taking single stranded DNA as template, described single stranded DNA is polyinosinic acid (PI) or poly (PC).

In a kind of above-mentioned method of synthesizing silver nanoclusters taking single stranded DNA as template, described reaction is carried out under condition of ice bath.

In a kind of above-mentioned method of synthesizing silver nanoclusters taking single stranded DNA as template, described cushioning liquid is the mixed solution of ammonium acetate and acetic acid composition. Can prevent the silver nanoclusters reunion forming.

In a kind of above-mentioned method of synthesizing silver nanoclusters taking single stranded DNA as template, the diameter of described silver nanoclusters is less than 5nm.

In a kind of above-mentioned method of synthesizing silver nanoclusters taking single stranded DNA as template, described reducing agent is sodium borohydride.

Compared with prior art, in the present invention, we have set up a kind of simple, low cost, green taking commercially available polyinosinic acid (PI) and poly (PC) as template, method is synthesized silver nanoclusters reliably. Fluorescence utilizes, and as polyinosinic acid and the synthetic fluorescence silver nanoparticle crystalline substance of cytidine monophosphate, does not need special design and pretreatment. This novel silver nanoclusters has good selective and sensitivity, can be applicable to detect Hg2+, be feasible and have developing prospect for monitoring pollutant in water.

Brief description of the drawings

Fig. 1. the structural formula of polyinosinic acid (A) and poly (B).

Fig. 2. the mechanism figure of synthetic AgNCs.

Fig. 3 .(A) the UV absorption figure of PI-AgNCs under the different reaction time (curve a-k:10,20,30,40,50,60,70, and 80 minutes); (B) the UV absorption figure (curve a-k:5,10,25,35,50,65,90,170,230,350, and1250 minute) of PC-AgNCs under the different reaction time; (C) PI-AgNCs(a) and PC-AgNCs(b) corresponding fluorescence spectrum is over time; Illustration: the uv absorption spectra while not adding NaBH4.

Fig. 4 .PI-AgNCs(A) and fluorescence excitation utilizing emitted light spectrogram PC-AgNCs(B).

Fig. 5 .PI-AgNCs(a) and fluorescence lifetime figure PC-AgNCs(b).

Fig. 6 .PI-AgNCs(A) and PC-AgNCs(B) at existence 0.7 μ molL-1Different metal ion (Hg2+,Ag+,Fe3+,Zn2+,Ca2+,Mn2+,Mg2+,Co2+,Pb2+,Ni2+,Cd2+,andCu2+Ion) relative fluorescence value time.

Fig. 7 .(A) add variable concentrations Hg2+The fluorescence emission spectrum (curve a-h) of rear PI-AgNCs: 0.133,0.267,0.400,0.533,0.667,0.800,0.933 and 1.07 μ M, illustration: fluorescence intensity and Hg2+The linear relationship of concentration; (B) add variable concentrations Hg2+The fluorescence emission spectrum (curve a-m) of rear PC-AgNCs: 0.667,1.33,2.00,2.67,3.33,4.00,4.67,5.33,6.00,6.67,7.33,8.00 and 8.67 μ M, illustration: fluorescence intensity and log[Hg2+] linear relationship.

Fig. 8. exist and do not have other metal ion (Hg2+5 times of concentration) situation under, add 0.7 μ MHg2+Time measure PI-AgNCs(A) and (I PC-AgNC(B)0-I)/I0Value.

Detailed description of the invention

Be below specific embodiments of the invention by reference to the accompanying drawings, technical scheme of the present invention is further described, but the present invention be not limited to these embodiment.

Embodiment 1

In the present embodiment, taking polyinosinic acid (PI) as template, synthetic fluorescence silver nanoclusters. Synthetic method is specific as follows: get respectively 0.2mL2.0%PI and 100 μ L10mmolL-1AgNO3, joins the cushioning liquid (0.09molL that 10mL is made up of ammonium acetate and acetic acid-1, pH=4.0) in. After mixed solution, make it at 0 DEG C, wait for 15 minutes, the 0.1mmolL then newly configuring to this mixed solution and dripping 20 μ L in the condition stirring-1Sodium borohydride solution. React after 15 minutes, the mixed solution that reaction is obtained is put dark place at room temperature 8 hours with stand-by again. In course of reaction, along with the amount of reducing agent increases, color burn, solution is faded to brown by light color, along with the carrying out of reaction, the Ag particle of harsh one-tenth is large, along with the increase of time, due to Ostwald ripening law, particle diminishes, thus the yellow nano-cluster forming.

Embodiment 2

In the present embodiment, taking poly (PC) as template, synthetic fluorescence silver nanoclusters.

Synthetic method is specific as follows: get respectively 0.5mL5.0%PI and 200 μ L12mmolL-1AgNO3, joins the cushioning liquid (0.08molL that 25mL is made up of ammonium acetate and acetic acid-1, pH=5.0) in. After mixed solution, make it at 0 DEG C, wait for 15 minutes, the 0.1mmolL then newly configuring to this mixed solution and dripping 50 μ L in the condition stirring-1Sodium borohydride solution. React after 15 minutes, the mixed solution that reaction is obtained is put dark place at room temperature 8 hours with stand-by again. In course of reaction, along with the amount of reducing agent increases, color burn, solution is faded to brown by light color, along with the carrying out of reaction, the Ag particle of harsh one-tenth is large, along with the increase of time, due to Ostwald ripening law, particle diminishes, thus the yellow nano-cluster forming.

In above-mentioned two embodiment, the same single stranded DNA as possessing cytimidine and guanine of polyinosinic acid (PI) and poly (PC), than bases adenine and thymidine, with Ag+There is stronger affinity. In addition the two keys of the ketonic oxygen existing at different rings, and nitrogen play key effect for preparing hyperfluorescenceZeng Yongminggaoyingguang silver nanoclusters. This is mainly owing to the resonances that form in single stranded DNA (as PI and PC). Specifically, the O in resonances-Trend towards and Ag+In conjunction with, form finely disseminated Ag+. Therefore, in conjunction with Ag+To carry out in-situ reducing along template. The key factor of another nanocluster formation is the secondary structure of single stranded DNA. In addition, template is vital for stablizing newly-generated silver nanoclusters and further producing fluorescence silver nanoparticle.

We use luminescent spectrum instrument to carry out fluoremetry to the product in above-mentioned two embodiment, and uv-visible absorption spectra is measured by UV/visible spectrophotometer, detect the forming process of silver nanoclusters with this.

For PI silver nanoclusters, there is no sodium borohydride, in ultraviolet-visible spectrum, do not observe absworption peak (illustration of Fig. 3 A). Add after sodium borohydride, absorption peak appears at 400nm place, and adsorption strength reduces (Fig. 3 A) along with the increase in reaction time. Meanwhile, with regard to PC silver nanoclusters, its absworption peak broadens gradually, and occurs obvious red shift (Fig. 3 B) at 405nm to 440nm. The time that further increases reaction, the adsorption peak of silver nanoclusters can split into two peaks (PI silver nanoclusters: 345nm and 430nm; PC silver nanoclusters: 395nm and 515nm).

PI silver nanoclusters is very similar with PC silver nanoclusters excitation and emission spectra spectrum separately. Increase in starting stage fluorescence intensity, reach after the plateau, then along with the further prolongation fluorescence intensity of time slowly decline (Fig. 3 C). With regard to by PI silver nanoclusters, occur hyperfluorescenceZeng Yongminggaoyingguang excitation spectrum at 545nm place, and emission maximum spectrum appears at 585nm place (Fig. 4 A). With regard to PC silver nanoclusters, excite accordingly with emission peak and lay respectively at 500nm and 575nm(Fig. 4 B). In a word, the synthetic silver nanoclusters of these two templates all has very strong fluorescence intensity, if but use polyadenylic acid (PA) is template, just do not have fluorescence signal to produce.

Fluorescence lifetime is one of characteristic parameter of fluorescence silver nanoclusters. PI silver nanoclusters (in Fig. 5 shown in a) and the fluorescence lifetime of PC silver nanoclusters (in Fig. 5 shown in b) be respectively 2.99ns and 1.19ns.

Utilize synthetic PI/PC silver nanoclusters to carry out Hg2+Detection, configuration 0.02mmolL-1Hg2+Solution is as detected solution, by the 0.02mmolL of different volumes-1Hg2+Solution joins in 3mLPI/PC silver nanoclusters solution, and solution is stablized 5 minutes after mixing, and then measures the fluorescence spectrum of PI/PC silver nanoclusters.

As shown in Figure 7, the fluorescence of AgNCs not only can be by Hg2+Quencher fast, and quencher intensity and the Hg of AgNCs2+Concentration is closely related, and the intensity of fluorescent quenching is along with Hg2+The increase of concentration and become more obvious. Fluorescence intensity and Hg2+Relation between concentration can be by Stern-Volmer equation I0/I=1+KSV[Q] analyzes and draws. The fluorescence intensity of PI-AgNCs is at Hg2+Concentration is 0.05 μ molL-1-1.0μmolL-1(R2=0.9747) in scope time, linearity weakens.

For PC-AgNCs, the weakening and Hg of fluorescence intensity2+The logarithm of concentration is linear. This processing was once respectively used to taking AgNCs as the little molecule (R of probe in detecting2=0.99) with taking AuNCs as probe (R2=0.96) detect Hg2+

In testing process, the fluorescence of AgNCs can be with micro-Hg2+Quencher fast within 5 minutes, this can measure Hg more quickly2+. In order to test AgNCs to Hg2+That measures is selective, the metal ion (Ag that other are relevant with environment+,Fe3+,Zn2+,Ca2+,Mn2+,Mg2+,Co2+,Pb2+,Ni2+,Cd2+And Cu2+Ion) be used under the same conditions test (Fig. 6). Apparently, change very littlely in the fluorescence intensity that adds PI-AgNCs before and after other metal ions or PC-AgNCs, this measures Hg with us2+Time situation difference. In other words, for measuring Hg2+Content, PI-AgNCs or PC-AgNCs are than better as the synthetic AgNCs performance of template with little molecule dihydrolipoic acid. Meanwhile, selective and selectively similar as the synthetic AgNCs of template with bovine serum albumin of our synthetic nano-cluster. The more important thing is, be different from a lot of organic fluorescence probes, our detection is at Cd2+And Pb2+Existence under to measure Hg2+Content do not disturb. These results prove PI-AgNCs or PC-AgNCs Hg in measurement biology and environmental samples2+Content aspect have very strong feasibility.

In conjunction with PI-AgNCs and PC-AgNCs, for measuring Hg2+The measurement category of content becomes wider. Taking PI-AgNCs as example, when signal to noise ratio is 3(S/N=3) time detect and be limited to 3.0nmolL-1(table 1), maximum permissible value (1ppb, the 5nmolL of this Mercury in Drinking Water content of announcing than European Union-1) low. For PC-AgNCs, detect and be limited to 9.0nmolL-1, maximum permissible value (2ppb, the 10nmolL of the Mercury in Drinking Water content of announcing lower than EPA-1). Synthetic nano-cluster with delivered in the past about Hg2+Sensor has been compared better sensitivity.

Table 1.PI-Ag and PC-AgNCs are detecting Hg2+The comparison of aspect of performance

aF, fluorescence intensity; C, Hg2+Concentration

In the quencher stage, O in resonances-With Hg2+In conjunction with having caused silver atoms to be extruded. Therefore, silver nanoclusters is destroyed, corresponding fluorescence signal cancellation subsequently. As expected, be used as polyadenylic acid (PA) for template, can't detect fluorescence signal, because there is no resonances in PA.

Interference while measuring mercury ion in order to test PI-AgNCs and PC-AgNCs, under other condition same cases, we have explored and have been five times in Hg2+Nine metal ion species (Ag of concentration+,Fe3+,Zn2+,Ca2+,Mn2+,Mg2+,Co2+,Pb2+And Ni2+Ion) to the disturbed condition (Fig. 8) of measuring. This nine metal ion species is to analyzing Hg2+Measure all noiseless. Therefore, prepared AgNCs shows good antijamming capability, and this is to Hg in the real-time detection of biological in scene and environmental samples2+Content is highly beneficial.

We have further inquired into PI-AgNCs and PC-AgNCs Hg in detection actual sample (the lake water sample of the new lunar lacus of Zhejiang Normal University)2+The practicality of content. This mixture mixes the Hg of variable concentrations2+Standard liquid. Hg in lake water sample2+Concentration has been listed (table 2). Clearly, be with PI-AgNCs and PC-AgNCs the Hg that probe in detecting goes out2+Concentration be almost identical and the rate of recovery of PI-AgNCs and PC-AgNCs within the scope of 95.3%-105.0%. In addition it is high when, the rate of recovery of mark-on water sample is made probe than with AuNCs. Meanwhile, the result of corresponding relative standard deviation (RSD) is satisfactory, and these results show that this method is feasibility.

The Hg that table 2. detects as probe taking PI-AgNCs or PC-AgNCs in actual water sample2+The parameters obtaining

Should be appreciated that, in claims of the present invention, description, all " comprising ... " is all interpreted as open implication, and namely its implication is equal to " at least containing ... ", and should not be construed as enclosed implication, its implication should not be construed " only comprising ... ".

Specific embodiment described herein is only to the explanation for example of the present invention's spirit. Those skilled in the art can make various amendments or supplement or adopt similar mode to substitute described specific embodiment, but can't depart from spirit of the present invention or surmount the defined scope of appended claims.

Claims (3)

1. a method for synthetic silver nanoclusters taking single stranded DNA as template, is characterized in that,The method be the single stranded DNA that possesses cytimidine and guanine be template, with Ag+Solution is mixedAfter closing, add after reducing agent reduces and make the silver nanoclusters with fluorescence, described single stranded DNAFor polyinosinic acid PI or poly PC, the concrete steps of described method comprise:
A, get a certain amount of single stranded DNA and Ag respectively+Solution, joins in cushioning liquidMix, after mixing, wait for that certain hour enters step b again;
B, in the mixed solution in step a, add reducing agent, reduction obtains water-soluble glimmeringLight silver nanoclusters product;
Reaction is carried out under condition of ice bath, and described cushioning liquid is ammonium acetate and vinegarThe mixed solution of acid composition.
2. one according to claim 1 is synthesized silver nanoparticle taking single stranded DNA as templateBunch method, it is characterized in that, the diameter of described silver nanoclusters is less than 5nm.
3. one according to claim 1 and 2 is synthetic silver-colored taking single stranded DNA as templateThe method of nano-cluster, is characterized in that, described reducing agent is sodium borohydride.
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