CN107884376A - Ratiometric fluorescent probe for mercury ion detecting and preparation method thereof - Google Patents
Ratiometric fluorescent probe for mercury ion detecting and preparation method thereof Download PDFInfo
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- CN107884376A CN107884376A CN201711168858.9A CN201711168858A CN107884376A CN 107884376 A CN107884376 A CN 107884376A CN 201711168858 A CN201711168858 A CN 201711168858A CN 107884376 A CN107884376 A CN 107884376A
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Abstract
The invention provides a kind of Ratiometric fluorescent probe of mercury ion detecting, double emission nanometer material gold/ag nano-clusters that probe is wrapped up by bovine serum albumin(BSA) are formed, and its fluorescence emission wavelengths is 400 550nm and 580 680nm.This pair transmitting gold/ag nano-cluster is for reducing agent and stabilizer with bovine serum albumin(BSA) (BSA), is synthesized using " treating different things alike " method.Gold/ag nano-cluster (Au/Ag NCs) of synthesis has two emission peaks, and wherein Ag NCs are as reference signal, and Au NCs are as response signal, for Hg2+Selective recognition.When the bimetal nano cluster is as Ratiometric fluorescent probe, Ag NCs fluorescence signal intensities are held essentially constant, and Au NCs can optionally with Hg2+With reference to so as to cause fluorescent quenching.This method is quick and easy, and test limit is low, and the range of linearity is wide, and selectivity is good.
Description
Technical field
The invention belongs to mercury ion detecting technical field, and in particular to a kind of Ratiometric fluorescent probe of mercury ion detecting.
Background technology
With the rapid development of economy, in environment heavy metal pollution getting worse, seriously endangered human health and ring
Border safety.Wherein mercury ion (Hg2+) not only pollute environment as a kind of heavy metal ion and also have significant damage to human body, it can lead
Hypoevolutism and some health problems are caused, such as damaged brain and nervous system, cause cognitive Bias, minamata disease, cause motion to hinder
Hinder and influence internal system etc..Simultaneously with mercury ion (Hg2+) for representative pollutant once enter environment after, will not quickly divide
Solution, but extended residual produces unfailing pollution in soil, water, air.Therefore find a kind of efficient, easy, quick
Mercury ion detecting method with regard to significant.The more ripe method of detection heavy metal ion is atom spectrum side at present
Method, including atomic absorption spectrum, atomic fluorescence spectrometry, atomic emission spectrometry and inductivity coupled plasma mass spectrometry, electrification
Credit analyse the methods of, but these methods in the prevalence of analytical procedure is cumbersome, sample pre-treatments are complicated, time-consuming, testing expense is high,
The shortcomings of high energy consumption.
XRF due to its analysis is sensitive and accurate, simple to operate, selectivity good, convenient and efficient and by extensively it is good
Comment, extensive concern is obtained in the field such as environmental monitoring and food security.In recent years, fluorescence metal nanocluster is widely adopted work
It is used for Hg for a kind of material of new replacement organic fluorescence group2+Detection., quantum production simple with modification is synthesized because it has
Rate is high, the advantages that water-soluble, photostability and good biocompatibility, therefore turns into preferable fluorescence probe.With monometallic nanometer
Cluster is compared, and bimetal nano cluster has a wide range of applications in catalysis, magnetics, optics and biological diagnosis etc., simultaneously because its
Special cooperative effect, has been to be concerned by more and more people.Existing mercury ion fluorescence probe generally uses single fluorescent quenching
Pattern, measurement result easily by environmental factor (temperature, pH, solvent polarity etc.), optical quenching, light source stability, detectable substance concentration and
The factors such as instrument condition change influence, and cause data distortion, so as to reduce the accuracy of measurement result.
Ratio fluorescent method is by measuring the fluorescence intensity at two different wave lengths, being determined using its ratio as signal parameter
The analysis method of object.This method can by measuring two different transmission signals simultaneously so as to provide built-in amendment,
Detected compared to single channel, it is possible to provide more accurate data analysis, can preferably avoid systematic error and human error, have
More preferable reappearance and stability.For the conventional fluorescent probe based on single fluorescence signal, it can reduce or eliminate inspection
Survey the data distortions caused by factor such as concentration of substrate, external environment condition and instrument condition change.Meanwhile two kinds of emitting at different wavelengths light
The change of line strength can cause the change of detection architecture color, make that detection process is relatively reliable, easy resolution, it is easier to the naked eye
Distinguish, be suitable for Visual retrieval.
Patent of invention CN201110409082.1 is disclosed a kind of highly sensitive detects trace mercury ion in water with high selectivity
Silver nanoclusters fluorescence probe, silver nanoclusters are made up of 1 to hundreds of silver atoms, and the size of silver nanoclusters is less than 3nm, and fluorescence is visited
Pin sends strong visible fluorescence at room temperature, and its launch wavelength scope is 400~800nm.Patent of invention CN200910218130.1
Disclosing a kind of fluorescent detection probe for mercury ions, probe is the nanogold particle of bovine serum albumin(BSA) functionalization, size is 1~
3nm, under 365nm light source, solution sends the light of red, tests ultraviolet spectra, in the case where 460~480nm wavelength excites,
630~655nm has very strong emission spectrum.And the gold or ag nano-cluster being manufactured separately only have single emission peak, it is impossible to
Component ratio type fluorescence probe.
Carbon point-gold nano cluster that patent of invention CN201510415104.3 discloses a kind of Visual retrieval mercury ion is double
Emission ratios type fluorescence probe, it is made up of double transmitting composite silicon dioxide nano particles；Double transmitting composite silicon dioxide nanoparticles
Son is answered using the silicon dioxide granule of carbon coated point by what covalent coupling gold nano cluster after kernel, its surface amination was formed
Close Nano particles of silicon dioxide.The material step of this method synthesis is very cumbersome, employs many organic reagents, and reaction is complicated,
And organic reagent is mostly harmful, environment is polluted.
The content of the invention
Problem be present for above-mentioned, it is an object of the invention to provide a kind of double transmittings for Visual retrieval mercury ion
Ratiometric fluorescent probe and preparation method thereof.The probe has single-minded selectivity and higher sensitivity to mercury ion, prepares
Method is simple, is adapted to amplification synthesis and practical application.
In order to realize foregoing invention purpose, the present invention adopts the following technical scheme that：
A kind of Ratiometric fluorescent probe of mercury ion detecting, double emission nanometer materials that probe is wrapped up by bovine serum albumin(BSA)
Gold/ag nano-cluster is formed, and its fluorescence emission wavelengths is 400-550nm and 580-680nm.
With bovine serum albumin(BSA) (BSA) for reducing agent and stabilizer in the nanocluster, gold/ag nano-cluster has two
Emission peak, wherein Ag NCs are as reference signal, and Au NCs are as response signal, for Hg2+Selective recognition.When this pair
When metal nanometer cluster is as Ratiometric fluorescent probe, Ag NCs fluorescence signal intensities are held essentially constant, and Au NCs can be selected
Selecting property and Hg2+With reference to so as to cause fluorescent quenching.The bimetal nano cluster can develop into Hg2+The ratio of content quantitative measure
Rate type-visualization fluorescence probe.
Preferably, gold/silver-colored mol ratio is 1 in the fluorescence probe:1, the cluster fluorescence synthesized under the mol ratio is substantially strong
In other mol ratios.
Preferably, the mass ratio of bovine serum albumin(BSA) and silver ion is 40-50 in the fluorescence probe：1.
The present invention for reducing agent and stabilizer, is synthesized with bovine serum albumin(BSA) (BSA) using " treating different things alike " method.Preparation side
Method is as follows：By certain density gold chloride (HAuCl4) solution and certain density silver nitrate (AgNO3) after solution is well mixed,
Bovine serum albumin solution is charged with again, a certain amount of NaoH solution is added after 2-10min, then in 20-50 DEG C of condition
Lower water-bath 10-15h, the solution for preparing gained are put into refrigerator preservation.
Preferably, the chlorauric acid solution concentration is 5mmol/L-25mmol/L, and silver nitrate solution concentration is 5mmol/L-
25mmol/L, and both mol ratios are 1:1.
Preferably, the concentration of bovine serum albumin solution and the NaoH solution is respectively 20-70mg/mL and 0.5-
5mol/L。
The BSA-Au/Ag NCs Ratiometric fluorescent probes detection mercury ion of the present invention：The dilution of mercury ion solution constant volume is taken to match somebody with somebody
A series of mercury standard liquids are made, by Hg2+Solution is added to BSA-Au/Ag NCs solution, then the constant volume under the conditions of cushioning liquid, mixing
Solution accelerates reaction to be thoroughly mixed with vortex concussion instrument at room temperature.Fluorescence emission spectrum is determined using XRF, and is turned
Move on to ultraviolet testing chamber and carry out Hg2+Visual retrieval, while in other ions such as Zn2+、Pb2+、Fe2+Body is investigated Deng in the presence of
The selectivity of system.The ratio between fluorescence intensity according to double emission peaks establishes standard curve, and contrast standard curve is obtained in testing sample
The concentration of mercury ion.
Preferably, the Hg2+Solution concentration is 20-200 μ g L-1, the mercury ion fluorescent in the concentration range, which is quenched, is in
Graded, it is convenient for Visual retrieval.
Preferably, 10,20,50,100,200 and 500 μ g L are prepared respectively-1Mercury ion standard liquid, by each concentration
Mercury ion solution is mixed with BSA-Au/Ag NCs solution with volume ratio 1: 1.Fluorescent quenching is obvious under the volume ratio, and being easy to can
Detected depending on changing.
Preferably, the pH value of phosphate buffer is 6.5-8.5, and the concentration of phosphate buffer is 10-50mmol/L.Phosphorus
The pH and concentration range of hydrochlorate are also the Optimal pH and concentration that optimization is drawn.
The beneficial effects of the present invention are：
1st, the present invention constructs the ratio based on gold/ag nano-cluster (Au/Ag NCs) of bovine serum albumin(BSA) parcel
Rate type fluorescent optical sensor is used for high sensitivity, the Visual retrieval Hg of high selectivity2+.A kind of be used for very specifically is invented
Gold/silver bimetal nanocluster fluorescence probe that mercury ion quick visualization detects in full pattern product and preparation method thereof.Prepare
Fluorescence probe can be used in Visual retrieval mercury ion, can Visual retrieval to 10 μ g L-1Mercury ion residue.This method detects
Limit relatively low, the range of linearity is wider, and available for the detection of actual sample, and high sensitivity, quick, simple, energy consumption is low, while this
It is that gold/silver bimetal nanocluster is used for Ratio-type and Visual retrieval mercury ion first, is quickly determined in environment heavy metal
In have boundless application prospect, this method is examined with being expected to be used for other metal nanometer cluster high selectivities and sensitivity
Check weighing metal ion.
2nd, the present invention constructs the gold/ag nano-cluster wrapped up with bovine serum albumin(BSA), component ratio type fluorescence probe, tool
There are two emission peaks, the data caused by factor such as detection substrate concentration, external environment condition and instrument condition change can be reduced or eliminated
Distortion etc..
3rd, the synthetic method of fluorescence probe of the present invention uses one-step synthesis method, and simply, green, environmental protection, energy consumption is low, nontoxic
Evil.
Brief description of the drawings
Fig. 1 is BSA-Au/Ag NCs ultraviolet spectra (a), fluorescence excitation spectrum (b) and fluorescence emission spectrogram of compound (c).
Fig. 2 is BSA-Au/Ag NCs shape appearance figures of the present invention.
Fig. 3 is BSA-Au/Ag NCs to various concentrations Hg2+Fluorescence response curve.470nm is Ag NCs fluorescence in figure
Emission peak, 630nm are Au NCs fluorescence emission peak.
Fig. 4 is fluorescence photos of the BSA-Au/Ag NCs of the present invention in the presence of various concentrations mercury ion.
Embodiment
The technical scheme in the embodiment of the present invention will be clearly and completely described below, it is clear that described implementation
Example only part of the embodiment of the present invention, rather than whole embodiments.It is common based on the embodiment in the present invention, this area
The every other embodiment that technical staff is obtained under the premise of creative work is not made, belong to the model that the present invention protects
Enclose.
Embodiment 1
A kind of Ratiometric fluorescent probe of mercury ion detecting, double emission nanometer materials that probe is wrapped up by bovine serum albumin(BSA)
Gold/ag nano-cluster is formed, and its fluorescence emission wavelengths is 400-550nm and 580-680nm.
Embodiment 2
A kind of Ratiometric fluorescent probe of mercury ion detecting, double emission nanometer materials that probe is wrapped up by bovine serum albumin(BSA)
Gold/ag nano-cluster is formed, and its fluorescence emission wavelengths is 400-550nm and 580-680nm.
Gold/silver-colored mol ratio is 1 in the fluorescence probe:1.
The mass ratio of bovine serum albumin(BSA) and silver ion is 40 in the fluorescence probe：1.
Embodiment 3
A kind of Ratiometric fluorescent probe of mercury ion detecting, double emission nanometer materials that probe is wrapped up by bovine serum albumin(BSA)
Gold/ag nano-cluster is formed, and its fluorescence emission wavelengths is 400-550nm and 580-680nm.
Gold/silver-colored mol ratio is 1 in the fluorescence probe:1.
The mass ratio of bovine serum albumin(BSA) and silver ion is 50 in the fluorescence probe：1.
Embodiment 4
A kind of Ratiometric fluorescent probe of mercury ion detecting, double emission nanometer materials that probe is wrapped up by bovine serum albumin(BSA)
Gold/ag nano-cluster is formed, and its fluorescence emission wavelengths is 400-550nm and 580-680nm.
Gold/silver-colored mol ratio is 1 in the fluorescence probe:1.
The mass ratio of bovine serum albumin(BSA) and silver ion is 45 in the fluorescence probe：1.
Embodiment 5
The preparation of the fluorescence probe of BSA-Au/Ag NCs Ratio-type mercury ions：Chlorauric acid solution and silver nitrate solution are mixed
After closing uniformly, then bovine serum albumin solution is charged with, NaoH solution, the then water-bath under the conditions of 20 DEG C are added after 2min
15h is reacted, produces fluorescence probe solution.
Embodiment 6
The preparation of the fluorescence probe of BSA-Au/Ag NCs Ratio-type mercury ions：Chlorauric acid solution and silver nitrate solution are mixed
After closing uniformly, then bovine serum albumin solution is charged with, NaoH solution is added after 10min, then the water under the conditions of 50 DEG C
Bath reaction 10h, produces fluorescence probe solution.
The chlorauric acid solution concentration is 5mmol/L, and silver nitrate solution concentration is 5mmol/L.
The concentration of bovine serum albumin solution and the NaoH solution is respectively 20mg/mL and 0.5mol/L.
Embodiment 7
The preparation of the fluorescence probe of BSA-Au/Ag NCs Ratio-type mercury ions：Chlorauric acid solution and silver nitrate solution are mixed
After closing uniformly, then bovine serum albumin solution is charged with, NaoH solution, the then water-bath under the conditions of 30 DEG C are added after 5min
12h is reacted, produces fluorescence probe solution.
The chlorauric acid solution concentration is 25mmol/L, and silver nitrate solution concentration is 25mmol/L.
The concentration of bovine serum albumin solution and the NaoH solution is respectively 70mg/mL and 5mol/L.
Embodiment 8
The preparation of the fluorescence probe of BSA-Au/Ag NCs Ratio-type mercury ions：Chlorauric acid solution and silver nitrate solution are mixed
After closing uniformly, then bovine serum albumin solution is charged with, NaoH solution, the then water-bath under the conditions of 30 DEG C are added after 5min
12h is reacted, produces fluorescence probe solution.
The chlorauric acid solution concentration is 15mmol/L, and silver nitrate solution concentration is 15mmol/L.
The concentration of bovine serum albumin solution and the NaoH solution is respectively 50mg/mL and 3mol/L.
Embodiment 9
The preparation of the fluorescence probe of BSA-Au/Ag NCs Ratio-type mercury ions, basic building-up process are as follows：
BSA-Au/Ag NCs synthesis：By 5mL, 10mmol/L chlorauric acid solution and 5mL, 10mmol/L silver nitrate
After solution is well mixed, then 5mL is charged with, 50mg/mL bovine serum albumin solution, 0.5mL is added after two minutes,
1mol/L NaoH solution, then the water-bath 12h under the conditions of 37 DEG C, the solution for preparing gained are put into ice under the conditions of 4 DEG C
Case preserves.Double compound BSA-Au/Ag NCs of transmitting UV-fluorescence spectrum and pattern is shown in Fig. 1 and Fig. 2.
Embodiment 10
The mercury ion detecting method of Ratiometric fluorescent probe of the present invention：The dilution of mercury ion solution constant volume is taken to prepare a series of mercury
Standard liquid, mercury ion solution is added in fluorescence probe solution, using phosphate buffer constant volume, mixed solution is used at room temperature
Vortex concussion instrument accelerates reaction complete, and fluorescence emission spectrum is determined using XRF, and is transferred to ultraviolet testing chamber progress
The Visual retrieval of mercury ion, standard curve is established according to the ratio between fluorescence intensity of double emission peaks, contrast standard curve is treated
The concentration of mercury ion in test sample product.
Embodiment 11
On the basis of embodiment 10：
10,20,50,100,200 and 500 μ g L are prepared respectively-1Mercury ion standard liquid, the mercury ion of each concentration is molten
Liquid is mixed with fluorescence probe solution with volume ratio 1: 1.
The pH value of phosphate buffer is 6.5, and the concentration of phosphate buffer is 10mmol/L.
Embodiment 12
On the basis of embodiment 10：
10,20,50,100,200 and 500 μ g L are prepared respectively-1Mercury ion standard liquid, the mercury ion of each concentration is molten
Liquid is mixed with fluorescence probe solution with volume ratio 1: 1.
The pH value of phosphate buffer is 8.5, and the concentration of phosphate buffer is 50mmol/L.
Embodiment 13
On the basis of embodiment 10：
10,20,50,100,200 and 500 μ g L are prepared respectively-1Mercury ion standard liquid, the mercury ion of each concentration is molten
Liquid is mixed with fluorescence probe solution with volume ratio 1: 1.
The pH value of phosphate buffer is 7, and the concentration of phosphate buffer is 30mmol/L.
Embodiment 14
The response condition that fluorescence probe determines to mercury ion
Take 1000mg/L mercury ion solution constant volume to dilute and prepare a series of mercury standard liquid (10,20,50,100,200,500 μ g
L-1).Take 100 μ L Hg2+Solution is added to 100 μ L BSA-Au/Ag NCs solution, then in phosphate buffer solution
(10mmol, pH 7) carries out being settled to 2 milliliters, and mixed solution accelerates reaction to be thoroughly mixed with vortex concussion instrument at room temperature.Should
Mixture is stirred at room temperature 20 minutes, and spectral measurement is then carried out on luminoscope, as a result sees Fig. 3.Cuvette is put into glimmering
Light spectrophotometer records fluorescence emission spectrum, and is transferred to ultraviolet testing chamber and carries out Hg in 365nm ultraviolet lights2+Visualization inspection
Survey, as a result see Fig. 4.Fluorescence intensities of the Ag NCs in 470nm is held essentially constant, and Au NCs red fluorescence intensities (630nm)
Gradually reduce；It is presented as that fluorescence intensity of the gold nanoclusters in 650nm reduces gradually from figure.It can be seen that by fluorescence photo, with
The increase of mercury ion content, the fluorescence of solution gradually become blueness by bright red, and fluorescence color change procedure is obvious, can be real
Existing Visual retrieval.
Fig. 3 shows：470nm is Ag NCs fluorescence emission peak in figure, and 630nm is Au NCs fluorescence emission peak.With
The increase of mercury ion content, 470nm fluorescence intensity is held essentially constant, and Au NCs red fluorescence intensities (630nm) are gradually
Reduce.
From fig. 4, it can be seen that with the increase of mercury ion content, the fluorescence color of BSA-Au/Ag NCs solution by red by
Fade to blueness, Visual retrieval can be achieved, can Visual retrieval to 10 μ g L-1Mercury ion residue.
Embodiment 15
The selectivity that fluorescence probe determines to mercury ion
100 μ g L are added under the same reaction conditions of embodiment 14-1Probe molecule, other metal ions are added, such as：
Zn2+,Mg2+,Ag+,Fe3+,Ni2+,Bi3+,K+,Co2+,Pb2+,Fe2+,Cd2+And Cu2+, concentration is 10000 μ g L-1.As a result send out
Now there was only mercury ion there occurs obvious fluorescent quenching, other metal ions without too significant change, therefore above-mentioned interference from
Under the conditions of existing for sub, probe still has preferable selectivity and sensitivity to mercury ion.
Embodiment 16
Detection of the fluorescence probe to actual sample
The Ratio-type of foundation-visualization mercury ion detecting new method is used for quick point of heavy metal ion in actual sample
Analysis.The methods of passing through mark-on reclaims detects to flesh of fish sample (DORM-4, Canada) and four kinds of rice samples into analysis has been carried out,
By calculating the rate of recovery, the stability of this method is investigated.As a result show, the rate of recovery of mercury is 95-98%'s in these samples
In the range of, this shows that the detection method has preferable accuracy and reliability.These results indicate that gold/the silver nanoparticle proposed
Cluster probe can delicately detect the Hg in actual sample2+。
A kind of double emission ratios of the gold for Visual retrieval mercury ion that the present invention is disclosed and proposed/silver bimetal nanocluster
Type fluorescence probe and preparation method, those skilled in the art can be by using for reference present disclosure, appropriate feed change and process route
Realized etc. link, although the method and technology of preparing of the present invention have been described by preferred embodiment, correlation technique people
Member can substantially not depart from present invention, methods and techniques described herein route is being modified in spirit and scope or again
Combination nova, to realize final technology of preparing.In particular, all similar replacements and change are to this area skill
It is it will be apparent that they are considered as being included in spiritual, scope and content of the invention for art personnel.
Claims (9)
- A kind of 1. Ratiometric fluorescent probe of mercury ion detecting, it is characterised in that double hairs that probe is wrapped up by bovine serum albumin(BSA) Penetrate nano material gold/ag nano-cluster to form, its fluorescence emission wavelengths is 400-550nm and 580-680nm.
- 2. the Ratiometric fluorescent probe of mercury ion detecting according to claim 1, it is characterised in that golden in the fluorescence probe/ Silver-colored mol ratio is 1:1.
- 3. the Ratiometric fluorescent probe of mercury ion detecting according to claim 1, it is characterised in that ox blood in the fluorescence probe The mass ratio of pure albumen and silver ion is 40-50：1.
- 4. according to the preparation method of Ratiometric fluorescent probe described in claim 1-3 any one, it is characterised in that by gold chloride After solution is well mixed with silver nitrate solution, then bovine serum albumin solution is charged with, it is molten that NaoH is added after 2-10min Liquid, then the water-bath 10-15h under the conditions of 20-50 DEG C, produces fluorescence probe solution.
- 5. the preparation method of Ratiometric fluorescent probe according to claim 4, it is characterised in that the chlorauric acid solution concentration For 5mmol/L-25mmol/L, silver nitrate solution concentration is 5mmol/L-25mmol/L, and both mol ratios are 1:1.
- 6. the preparation method of mercury Ratiometric fluorescent probe according to claim 4, it is characterised in that the bovine serum albumin(BSA) The concentration of solution and NaoH solution is respectively 20-70mg/mL and 0.5-5mol/L.
- 7. according to the mercury ion detecting method of Ratiometric fluorescent probe described in claim 1-3 any one, it is characterised in that take A series of mercury standard liquids are prepared in the dilution of mercury ion solution constant volume, mercury ion solution are added in fluorescence probe solution, using phosphoric acid Salt buffer constant volume, mixed solution accelerate reaction complete with vortex concussion instrument at room temperature, and fluorescence is determined using XRF Emission spectrum, and the Visual retrieval that ultraviolet testing chamber carries out mercury ion is transferred to, according to the ratio between fluorescence intensity of double emission peaks Standard curve is established, contrast standard curve obtains the concentration of mercury ion in testing sample.
- 8. the mercury ion detecting method of Ratiometric fluorescent probe according to claim 7, it is characterised in that 10 are prepared respectively, 20,50,100,200 and 500 μ g L-1Mercury ion standard liquid, by the mercury ion solution of each concentration and fluorescence probe solution with Volume ratio 1: 1 mixes.
- 9. the mercury ion detecting method of Ratiometric fluorescent probe according to claim 7, it is characterised in that phosphate buffer PH value be 6.5-8.5, the concentration of phosphate buffer is 10-50mmol/L.
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