CN107655868A - A kind of preparation method of water-soluble gold nano cluster and its application that copper trace in Chinese medicine is determined as fluorescence probe - Google Patents
A kind of preparation method of water-soluble gold nano cluster and its application that copper trace in Chinese medicine is determined as fluorescence probe Download PDFInfo
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Abstract
The application of copper trace in Chinese medicine is determined the invention discloses a kind of preparation method of water-soluble gold nano cluster and its as fluorescence probe, belongs to fluorescence analysis field.Technical scheme main points are：A kind of preparation method of water-soluble gold nano cluster, the steps such as the main preparation including Cefotaxime Sodium degradation product solution, the preparation of gold solution and the preparation of water-soluble gold nano cluster, and specifically disclose the application that obtained water-soluble gold nano cluster determines copper trace in Chinese medicine as fluorescence probe.The synthesis condition of the water-soluble gold nano cluster of the present invention is simply controllable, and obtained water-soluble gold nano cluster can for a long time store and have the advantages that good light stability, excitation wavelength range are wide and emission spectrum peak type is symmetrical；The Copper Ion method that the water-soluble gold nano cluster is established as copper ion fluorescence probe, there is high sensitivity, selective strong and strong antijamming capability, trace Cupper determination in Chinese medicine can be successfully applied to.
Description
Technical field
The invention belongs to fluorescence analysis field, and in particular to a kind of preparation method and its work of water-soluble gold nano cluster
The application of copper trace in Chinese medicine is determined for fluorescence probe.
Background technology
Copper is most important to health, can produce substantial amounts of enzyme with some protein bindings.But if intake copper mistake
Amount, it can react to form reactive oxygen free radical with molecular oxygen, can also destroy protein, nucleic acid and lipid.Copper is dived to human body
Toxicity and to caused by environment pollution have become countries in the world concern the problem of.Therefore, accurate and sensitive inspection is established
The method tool for surveying copper ion is of great significance.
Gold nanoclusters（AuNCs）It is a kind of new fluorescent nano material, is generally made up of several to dozens of gold atoms,
Size is less than 2nm, it has also become the area research such as material science and life science and using focus, they have fluorescence intensity it is big,
The advantages that toxicity is low, water-soluble good and biocompatibility is high, has in fields such as analysis detection, biomarker and fluorescence imagings
It is widely applied prospect.
The content of the invention
Present invention solves the technical problem that there is provided a kind of preparation method of water-soluble gold nano cluster and its as fluorescence
The application of copper trace in probe assay Chinese medicine, this method prepare water soluble fluorescence gold nano using the catabolite of Cefotaxime Sodium
Cluster（DCTX-AuNCs）, and based on copper ion there is selective quenching effect to establish one kind quickly to DCTX-AuNCs fluorescence
And easy high selection and the method for highly sensitive detection copper trace.
The present invention is to solve above-mentioned technical problem to adopt the following technical scheme that, a kind of preparation side of water-soluble gold nano cluster
Method, it is characterised in that concretely comprise the following steps：2.5mL is added in the CTX sodium solution that 5mL molar concentrations are 50mmol/L to rub
The sodium hydroxide solution that your concentration is 2mol/L, adds 20mL deionized waters, and room is cooled to cold water after 100 DEG C of reaction 1h
Temperature, adds hydrochloric acid solution and adjusts to neutrality and obtain Cefotaxime Sodium degradation product solution for standby；By 1g HAuCl4 .4H2O is dissolved in
In ionized water and it is settled to 500mL to obtain gold solution standby；14mL gold solutions are placed in reaction vessel and oil bath heating is extremely boiled
Rise, seethe with excitement 10min after add 7mL Cefotaxime Sodium degradation product solution, continue seethe with excitement 4h, treat solution by brown to purple powder again to
Stop heating when light yellow, naturally cool to room temperature, be then filtrated to get with 0.45 μm of hydrophilic PTFE syringe filters pale yellow
Color clear solution is water-soluble gold nano cluster, is placed in dark place in 4 DEG C of refrigerators and preserves, standby.
Water-soluble gold nano cluster of the present invention determines the application of copper trace in Chinese medicine as fluorescence probe, and its feature exists
It is in detailed process：
（1）The drafting of standard curve, the water-soluble gold nano cluster that 300 μ L are synthesized is taken in 5mL colorimetric cylinders, add concentration known ladder
The copper ion solution of degree, the phosphate buffer solution of 200 μ L pH=7 is added, 4mL is settled to ultra-pure water, in 30 DEG C of placements
40min, in incident and exit slit width it is 10nm with 1cm*1cm quartz sample pool, voltage is surveyed under conditions of being 850V
Measure fluorescence intensity, when copper ion concentration scope is 0.01-40 μm of ol/L, F0/ F and copper ion are in good linear relationship, are returned
The equation is returned to be：F0/ F=0.06968C+1.05944, coefficient R2=0.99425, detection is limited to 0.009 μm of ol/L, and copper ion is dense
Spend for 20 μm of ol/L parallel determinations 12 times, relative standard deviation 1.20%；
（2）Trace Cupper determination in Chinese medicine, Chinese medicine is washed with deionized water only, in 90 DEG C of drying, takes out and 200 mesh sieves is crossed after crushing
And in being saved backup in drier, accurately weigh above-mentioned traditional Chinese medicine sample 0.5g and be placed in beaker, 10mL salpeter solutions are added, are covered
Surface plate is stood overnight, and next day adds 10mL salpeter solutions and perchloric acid solution, is placed on electric hot plate and is heated to seething with excitement, and is kept molten
Liquid is under fluidized state until after solution clarification, is continued to be heated to emit white cigarette until white cigarette disperses, solution become colorless it is transparent,
Ultrapure water surface plate and beaker inwall are used after cooling, is transferred in 25mL volumetric flasks and is settled to 25mL with ultra-pure water and obtain
Traditional Chinese medicine sample solution for standby, the water-soluble gold nano cluster that 300 μ L are synthesized is taken in 5mL colorimetric cylinders, add traditional Chinese medicine sample solution, then
The phosphate buffer solution of 200 μ L pH=7 is added, 4mL is settled to ultra-pure water, 40min is placed in 30 DEG C, with 1cm*1cm's
Quartz sample pool is 10nm in incident and exit slit width, and voltage measures fluorescence intensity, Ran Hougen under conditions of being 850V
According to regression equation F0/ F=0.06968C+1.05944 obtains the concentration of copper ion in traditional Chinese medicine sample solution, and according to traditional Chinese medicine sample
Mass Calculation obtain the content of copper trace in Chinese medicine.
Further preferably, the Chinese medicine is radix glycyrrhizae, Poria cocos, rhizoma alismatis or the red sage root.
The present invention has the advantages that compared with prior art：The synthesis condition of the water-soluble gold nano cluster of the present invention
Simple controllable, obtained water-soluble gold nano cluster can for a long time store and have good light stability, excitation wavelength range wide and launch
The advantages that spectrum peak type is symmetrical, there is preferable fluorescence property, water solubility gold in pH=2-7, higher ionic strength range
The Copper Ion method that nano-cluster is established as copper ion fluorescence probe, there is high sensitivity, the strong and anti-interference energy of selectivity
The features such as power is strong, trace Cupper determination in Chinese medicine can be successfully applied to.
Brief description of the drawings
Fig. 1 is exciting and launching fluorescence pattern for water-soluble gold nano cluster；
Fig. 2 is water-soluble gold nano cluster and DCTX UV, visible light extinction spectrum figure；
Fig. 3 is pH to water-soluble gold nano cluster stability influence block diagram；
Fig. 4 is ionic strength to water-soluble gold nano cluster stability influence block diagram；
Fig. 5 is the time to water-soluble gold nano cluster stability influence block diagram；
Fig. 6 is the light stability curve of water-soluble gold nano cluster；
Fig. 7 is the selection performance curve of water-soluble gold nano cluster assay method；
Fig. 8 is the interference experiment result block diagram of water-soluble gold nano cluster assay method.
Embodiment
The above of the present invention is described in further details by the following examples, but this should not be interpreted as to this
The scope for inventing above-mentioned theme is only limitted to following embodiment, and all technologies realized based on the above of the present invention belong to this hair
Bright scope.
Embodiment 1
1st, the preparation of water-soluble gold nano cluster：
Cefotaxime Sodium degradation product（DCTX）The preparation of solution
The hydrogen-oxygen that 2.5mL molar concentrations are 2mol/L is added in the CTX sodium solution that 5mL molar concentrations are 50mmol/L
Change sodium solution, add 20mL deionized waters, room temperature is cooled to cold water after 100 DEG C of reaction 1h, add hydrochloric acid solution tune
Section to neutrality obtains Cefotaxime Sodium degradation product solution for standby.
The preparation of gold solution
By 1g HAuCl4 .4H2O is dissolved in deionized water and is settled to 500mL to obtain gold solution standby.
The preparation of water-soluble gold nano cluster
14mL gold solutions are placed in reaction vessel and oil bath heating is to seething with excitement, 7mL Cefotaxime Sodiums drop is added after the 10min that seethes with excitement
Solve thing solution, continue seethe with excitement 4h, when solution by brown to purple powder again to it is light yellow when stopping heating, naturally cool to room temperature,
Then shallow yellow transparent solution i.e. water-soluble gold nano cluster is filtrated to get with 0.45 μm of hydrophilic PTFE syringe filters, is placed in 4
Dark place preserves in DEG C refrigerator, standby.
2nd, characterize：
Fluorescence spectrum
Fig. 1 is shown in exciting and launching fluorescence pattern for water-soluble gold nano cluster, and its excitation wavelength is 321nm, and launch wavelength is
420nm, and fluorescence emission spectrogram of compound peak type is symmetrical, while there is very wide fluorescence exciting wavelength scope（250-400nm）, table
Bright obtained water-soluble gold cluster has preferable fluorescence property.
Ultraviolet spectra
Water-soluble gold nano cluster and DCTX UV, visible light extinction spectrum are shown in that Fig. 2, DCTX have very little in 250nm and 300nm or so
Absworption peak, and water-soluble gold nano cluster is almost without obvious surface plasmon peak, and this is mainly due to working as golden nanometer particle
Particle diameter 2nm or so when, cause its surface plasma absworption peak to drastically reduce due to quantum size effect, peak shape broadens even
Disappear, thus provable water-soluble gold nano cluster successfully synthesizes.
3rd, the stability of water-soluble gold nano cluster：
Influences of the pH to water-soluble gold nano cluster stability
Fig. 3 gives pH to water-soluble gold nano cluster stability influence block diagram.As seen from the figure：It is glimmering in the range of pH=2-7
Luminous intensity is held essentially constant, and the water-soluble gold nano cluster of this explanation synthesis maintains very well in the range of wider pH=2-7
Stability.
Influence of the ionic strength to water-soluble gold nano cluster stability
Ionic strength increases to 200mmol/L from 5mmol/L, as shown in figure 4, ionic strength is to water-soluble gold nano stability
Have little to no effect.
Influence of the time to water-soluble gold nano cluster stability
As shown in figure 5, the water-soluble gold nano cluster of synthesis fluorescence intensity level within the time of 2 months does not have downward trend, this says
Bright water-soluble gold nano cluster is also unaffected through storing its stability for a long time.
The optical stability of water-soluble gold nano cluster
As shown in fig. 6, continuous radiation 3h, relative to initial fluorescent intensity, the fluorescence intensity of water-soluble gold nano cluster remains 71%
（Launch wavelength is 321nm）, the results showed that water-soluble gold nano cluster has preferable photostability.
4th, the application of water-soluble gold nano cluster：
The drafting of standard curve
The water-soluble gold nano cluster that 300 μ L are synthesized is taken in 5mL colorimetric cylinders, adds the copper ion solution of concentration known gradient, then
The phosphate buffer solution of 200 μ L pH=7 is added, 4mL is settled to ultra-pure water, 40min is placed in 30 DEG C, with 1cm*1cm's
Quartz sample pool is 10nm in incident and exit slit width, and voltage measures fluorescence intensity under conditions of being 850V, copper from
When sub- concentration range is 0.01-40 μm of ol/L, F0/ F and copper ion are in good linear relationship, and regression equation is：F0/F=
0.06968C+1.05944, coefficient R2=0.99425, detection is limited to 0.009 μm of ol/L, and copper ion concentration is 20 μm of ol/L
Parallel determination 12 times, relative standard deviation 1.20%.
The selectivity of assay method is strong
The water-soluble gold nano cluster that 300 μ L are synthesized is taken to be separately added into Cu in 5mL colorimetric cylinders2+、Zn2+、Al3+、Ni2+、Co2+、
Pb2+、Cr3+、Cd2+、Mn2+、Mg2+、K+、Bi3+、Ba2+、Ca2+、Fe3+And Hg2+Solution, final concentration of metal ions be 20 μM,
Blank is not added with any ion, adds the phosphate buffer solution of 200 μ L pH=7, is settled to 4mL with ultra-pure water, is put in 30 DEG C
Put 40min.As shown in fig. 7, the addition of copper ion, the fluorescent quenching of system is maximum, and other metals are to the fluorescence intensity of system
Influence very small.
The strong antijamming capability of assay method
As shown in figure 8, the Zn of equimolar concentration2+、Al3+、Ni2+、Co2+、Pb2+、Cr3+、Cd2+、Mn2+、Mg2+、K+、Bi3+、Ba2+、
Ca2+、Fe3+And Hg2+Deng the presence of metal ion, the measure of copper ion is not almost interfered with.
Application in Chinese medicine copper trace measure
By Chinese medicine（Radix glycyrrhizae, Poria cocos, rhizoma alismatis or the red sage root）It is washed with deionized water only, in 90 DEG C of drying, takes out and 200 mesh sieves are crossed after crushing
And in being saved backup in drier, accurately weigh above-mentioned traditional Chinese medicine sample 0.5g and be placed in beaker, 10mL salpeter solutions are added, are covered
Surface plate is stood overnight, and next day adds 10mL salpeter solutions and perchloric acid solution, is placed on electric hot plate and is heated to seething with excitement, and is kept molten
Liquid is under fluidized state until after solution clarification, is continued to be heated to emit white cigarette until white cigarette disperses, solution become colorless it is transparent,
Ultrapure water surface plate and beaker inwall are used after cooling, is transferred in 25mL volumetric flasks and is settled to 25mL with ultra-pure water and obtain
Traditional Chinese medicine sample solution for standby, the water-soluble gold nano cluster that 300 μ L are synthesized is taken in 5mL colorimetric cylinders, add traditional Chinese medicine sample solution, then
The phosphate buffer solution of 200 μ L pH=7 is added, 4mL is settled to ultra-pure water, 40min is placed in 30 DEG C, with 1cm*1cm's
Quartz sample pool is 10nm in incident and exit slit width, and voltage measures fluorescence intensity, Ran Hougen under conditions of being 850V
According to regression equation F0/ F=0.06968C+1.05944 obtains the concentration of copper ion in traditional Chinese medicine sample solution, and according to traditional Chinese medicine sample
Mass Calculation obtain the content of copper trace in Chinese medicine.
It is measured respectively with fluorescence method proposed by the present invention and atomic absorption method, the results are shown in Table 1, every group of experiment repeats 5
It is secondary.T is examined（t0.05,4=2.78）As a result show, fluorescence method proposed by the present invention is used successfully to trace Cupper determination in Chinese medicine, accurate
Really reliable, method is feasible.
The content of copper trace in the water-soluble gold nano cluster fluorescence probe method of table 1 measure Chinese medicine
Claims (4)
1. a kind of preparation method of water-soluble gold nano cluster, it is characterised in that concretely comprise the following steps：It is in 5mL molar concentrations
The sodium hydroxide solution that 2.5mL molar concentrations are 2mol/L is added in 50mmol/L CTX sodium solution, adds 20mL
Deionized water, with cold water room temperature is cooled to after reacting 1h in 100 DEG C, adds hydrochloric acid solution and adjust to neutrality and obtain CTX
Sodium degradation product solution for standby；By 1g HAuCl4 .4H2O is dissolved in deionized water and is settled to 500mL to obtain gold solution standby；Will
14mL gold solutions are placed in reaction vessel and oil bath heating is to seething with excitement, and it is molten to add 7mL Cefotaxime Sodium degradation products after the 10min that seethes with excitement
Liquid, continue seethe with excitement 4h, when solution by brown to purple powder again to it is light yellow when stopping heating, naturally cool to room temperature, Ran Houyong
0.45 μm of hydrophilic PTFE syringe filters are filtrated to get shallow yellow transparent solution i.e. water-soluble gold nano cluster, are placed in 4 DEG C of refrigerators
Middle dark place preserves, standby.
2. water-soluble gold nano cluster made from the method described in claim 1 as fluorescence probe determine Chinese medicine in copper trace should
With.
3. water-soluble gold nano cluster according to claim 2 determines the application of copper trace in Chinese medicine as fluorescence probe, its
It is characterised by that detailed process is：
（1）The drafting of standard curve, the water-soluble gold nano cluster that 300 μ L are synthesized is taken in 5mL colorimetric cylinders, add concentration known ladder
The copper ion solution of degree, the phosphate buffer solution of 200 μ L pH=7 is added, 4mL is settled to ultra-pure water, in 30 DEG C of placements
40min, in incident and exit slit width it is 10nm with 1cm*1cm quartz sample pool, voltage is surveyed under conditions of being 850V
Measure fluorescence intensity, when copper ion concentration scope is 0.01-40 μm of ol/L, F0/ F and copper ion are in good linear relationship, are returned
The equation is returned to be：F0/ F=0.06968C+1.05944, coefficient R2=0.99425, detection is limited to 0.009 μm of ol/L, and copper ion is dense
Spend for 20 μm of ol/L parallel determinations 12 times, relative standard deviation 1.20%；
（2）Trace Cupper determination in Chinese medicine, Chinese medicine is washed with deionized water only, in 90 DEG C of drying, takes out and 200 mesh sieves is crossed after crushing
And in being saved backup in drier, accurately weigh above-mentioned traditional Chinese medicine sample 0.5g and be placed in beaker, 10mL salpeter solutions are added, are covered
Surface plate is stood overnight, and next day adds 10mL salpeter solutions and perchloric acid solution, is placed on electric hot plate and is heated to seething with excitement, and is kept molten
Liquid is under fluidized state until after solution clarification, is continued to be heated to emit white cigarette until white cigarette disperses, solution become colorless it is transparent,
Ultrapure water surface plate and beaker inwall are used after cooling, is transferred in 25mL volumetric flasks and is settled to 25mL with ultra-pure water and obtain
Traditional Chinese medicine sample solution for standby, the water-soluble gold nano cluster that 300 μ L are synthesized is taken in 5mL colorimetric cylinders, add traditional Chinese medicine sample solution, then
The phosphate buffer solution of 200 μ L pH=7 is added, 4mL is settled to ultra-pure water, 40min is placed in 30 DEG C, with 1cm*1cm's
Quartz sample pool is 10nm in incident and exit slit width, and voltage measures fluorescence intensity, Ran Hougen under conditions of being 850V
According to regression equation F0/ F=0.06968C+1.05944 obtains the concentration of copper ion in traditional Chinese medicine sample solution, and according to traditional Chinese medicine sample
Mass Calculation obtain the content of copper trace in Chinese medicine.
4. water-soluble gold nano cluster according to claim 3 determines the application of copper trace in Chinese medicine as fluorescence probe, its
It is characterised by：The Chinese medicine is radix glycyrrhizae, Poria cocos, rhizoma alismatis or the red sage root.
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| CN109655609A (en) * | 2019-02-20 | 2019-04-19 | 军事科学院军事医学研究院军事兽医研究所 | Platinum-nano flower and its preparation method and application |
| CN110530831A (en) * | 2019-08-26 | 2019-12-03 | 河南师范大学 | Fluorescence enhancement system and its application based on gold nanoclusters AuNCs@APAP |
| CN110530832A (en) * | 2019-08-26 | 2019-12-03 | 河南师范大学 | The method for selectively measuring 2,4- dinitrophenol dinitrophenolate in earth's surface water sample based on fluorescence analysis |
| CN110564403A (en) * | 2019-08-26 | 2019-12-13 | 河南师范大学 | Preparation method and application of dopamine-functionalized gold nanocluster |
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| CN115161018A (en) * | 2022-07-14 | 2022-10-11 | 河南师范大学 | Sulfur quantum dot fluorescent probe, preparation method thereof and application thereof in gold ion detection |
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