CN103424388B - A kind of TNT colorimetric fluorescent detection probe and application process thereof - Google Patents
A kind of TNT colorimetric fluorescent detection probe and application process thereof Download PDFInfo
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Abstract
The invention belongs to nanometer detection technical field, it is specifically related to a kind of 2,4, the colorimetric of 6-trinitrotoluene (TNT) and fluorescent dual-function detect probe and application process thereof, the present invention provides the gold nanoclusters of bovine serum albumin functionalization as colorimetric and double check probe, BSA-GNPs solution after dilution is taken as nanometer detection probe, is the amount hybrid reaction of 1:1 by volume with sample to be measured, observe the color change of TNT。Also filter paper can be cut into strip, be immersed in the BSA-GNPs aqueous solution of dilution 10 times, after taking-up, filter paper is dried, apply as Test paper。It is also used as colorimetric detection probes, the existence of TNT can be gone out under common sunshine condition by colorimetric determination。Can realizing original position quickly to detect, detection limit is low, can be applicable to more many condition and scope, simple to operate, cost is low, agents useful for same and operating process have no side effect。
Description
Technical field
The invention belongs to nanometer detection technical field, the colorimetric and the fluorescent dual-function that are specifically related to a kind of 2，4，6-trinitrotoluene (TNT) detect probe and application process thereof。
Background technology
2，4，6-trinitrotoluene (Trinitrotoluene, TNT, 2,4,6-trinitromethylbenzene) is a kind of faint yellow aromatic hydrocarbons crystal, and molten point is 354K, and it is with explosivity, is one of conventional Explosive ingredients。The trinitrotoluene of refine is very fixed。Different from nitroglycerin, it is all insensitive for friction, vibrations etc.。But it is with alkali reaction kickback, generate unstable compound。These compounds are all very sensitive to heat and shock。The toxicity of TNT is moderate toxicity, can percutaneous, respiratory tract, digestive tract invade, and main harm is chronic poisoning, and people is exposed to trinitrotoluene for a long time can be increased and suffer from anemia and the abnormal chance of liver function；Inject or sucked the animal also discovery of trinitrotoluene to affect blood and liver, spleen and send out big with other about immune bad influences；Also demonstrate the TNT reproductive function to male on evidence and have bad influence, and TNT is also listed in one and is potentially carcinogenic thing。Feed TNT can make urine blackening。Some military proving ground is polluted by TNT。Sewage produced by munitions can pollute the surface water and subsoil water。These contaminated water meeting pinkiness, this is because water is polluted by TNT。These pollutant are referred to as pink water, will clear up it very difficult and expensive。
Therefore, in recent years, pay close attention to widely and fruitful exploration the detection of ultra trace nitroaromatic and the exploration of relevant sensor array have been caused mechanism of social research。The laboratory detection of specific nitryl aromatic race explosive and vapor signal thereof is carried out widely already by the method for gas chromatograph-mass spectrometer, ion mobility spectrometry and neutron activity analysis etc.。These conventional analytical techniques disclosure satisfy that the basic demand in analysis, such as selectivity, reliability, accuracy and repeatability, but these detection methods are expensive, consuming time and loaded down with trivial details heavinesses, because in detection, sample must be an off detection scene and be sent to laboratory for analysis, it is impossible to enough accomplish the detection of real-time on-site。In sum, it is necessary to find a kind of method that can quickly and easily detect TNT。
For problem above, we have researched and developed a kind of method that gold nanoclusters utilizing functionalization detects TNT with the double function probe for colorimetric and fluoroscopic examination。The present invention provides the double function probe of a kind of colorimetric and fluoroscopic examination for detecting TNT, and provides its method simple to operate, Quantitative detection TNT。
Summary of the invention
It is an object of the invention to provide a kind of colorimetric detecting explosive TNT and fluorescent dual-function detection probe。
It is a further object of the present invention to provide the application process of a kind of detection probe detecting explosive TNT。
Fluorescent detection probe for mercury ions of the present invention is the fluorogold nano-cluster of bovine serum albumin (BSA) functionalization。
Bovine serum albumin molecular surface is rich in each seed amino acid, with the nanogold particle rich surface of bovine serum albumin functionalization containing amino and carboxylic group, the nitryl aromatic explosive TNT having the aromatic rings of electron deficiency is the acceptor of an electronics, thus demonstrate the fluorescent material surface to electron rich and have significantly high affinity, this luminescence generated by light passes through the electron transfer two direct cancellation of complex mechanism between electron acceptor and donor, what this cancellation depended on nitryl aromatic compound accepts electronic capability, therefore, pass through fluorescence detection method, can detect that the TNT of the trace contained in sample。
One, the synthesis of the fluorogold nano-cluster (BSA-GNanoclusters) of fluorescent probe bovine serum albumin functionalization:
The Nano-Au probe of bovine serum albumin (BSA) functionalization is [XieJP prepared by the method according to bibliographical information, ZhengYG, YingJY, Protein-DirectedSynthesisofHighlyFluorescentGoldNanoclus ters [J] .J.Am.Chem.Soc.2009,131 (3): 888 889.]。Concrete synthetic method is as follows: gold chloride is made into the stock solution of 10mM, under the water bath condition of 30～40 ° of C, it is that 1.5～3:1 is made into aqueous solution by the mass ratio of bovine serum albumin Yu two kinds of reactants of gold chloride, two kinds of solution mix under magnetic stirring, after 2～5 minutes, the NaOH solution adding the 1M accounting for mixed liquor volume 5% regulates the pH value of mixed solution, then under 30～40 ° of C water bath condition of constant temperature continuously stirred 10～24 hours。The color of solution is become dark-brown by the glassy yellow of original chlorauric acid solution。Fluorogold nano-cluster solution presents strong red emission under ultraviolet source, excites under the wavelength of 250～450nm, and fluorogold nano-cluster has strong emission spectrum at 630～660nm。
Two, detection TNT
Detection process: fluorogold nano-cluster (hereinafter referred to as the fluorogold nano-cluster) solution of the bovine serum albumin functionalization prepared has very strong fluorescent emission, in order to make the sensitivity in detection process improve, original solution is diluted 10～20 times, and the fluorogold nano-cluster solution after the dilution obtained is taken as nanometer detection probe。The gold nanoclusters detection probe of bovine serum albumin functionalization not only can indicate the existence of target detection thing TNT by the change of fluorescence, it is also used as colorimetric detection probes, do not need uviol lamp to excite, the existence of TNT can be gone out under common sunshine condition by colorimetric determination。In the process of fluorogold nano-cluster detection probe in detecting explosive, sample to be measured detects the amount that probe is 1:1 by volume and mixes with fluorogold nanocluster fluorescence, namely the solution example of the explosive of 100 μ L adds in 100 μ L nanometer detection probes, all of sensitivity experiment and selectivity experiment all carry out in above ratio, and specific experiment process is as follows:
1, detection sensitivity
When the fluorogold nano-cluster using bovine serum albumin functionalization detects probe as nanometer detection probe in detecting TNT, there is significantly high sensitivity。
In the experiment of inspection fluorogold nano-cluster detection probe sensitivity, taking concentration respectively is 10-3、5×10-4、10-4、5×10-5、10-5、5×10-6、10-6、5×10-7、10-7The fluorogold nano-cluster aqueous solution of the dilution 10 times of the TNT acetone soln 100 μ L of M and equivalent is reacted, it can be observed that 10-3、5×10-4、10-4、5×10-5The color generation significant change of the TNT of M concentration, color is from the original colourless redness that becomes, and the concentration along with TNT of the color of solution raises and deepens；Meanwhile, under the irradiation of uviol lamp, die down successively with the rising fluorescence intensity of TNT concentration。By ultraviolet spectra and fluorescence spectrum, it is possible to verify the sensitivity (accompanying drawing 4, accompanying drawing 5) of probe further。Therefore, nanometer detection probe can be used for detecting TNT as colorimetric and fluorescent dual-function probe。
2, selectivity
In the fluorogold nano-cluster namo fluorescence probe detection TNT of inspection bovine serum albumin functionalization optionally tests, by paranitrophenol, meta-nitrotoluene, picric acid, 2,4-dinitrotoluene (DNT), the toluene explosive similar to TNT structure with Nitrobenzol, as detection object, investigates the selectivity of detection probe。Taking concentration respectively is 10-3The TNT of M, paranitrophenol, meta-nitrotoluene, picric acid, 2, the acetone soln 100 μ L and 100 μ L of 4-dinitrotoluene (DNT) dilutes the fluorogold nano-cluster solution reaction of 10～20 times, can see that the fluorogold nano-cluster solution colour adding TNT changes, become redness, and other solution colours become yellow except paranitrophenol except without significant change。When irradiating under ultraviolet source, the fluorescence intensity of TNT dies down, and the fluorescence intensity of other solution does not have significant change, therefore, it is possible to judge that go out the namo fluorescence probe detection to TNT have good selectivity。
3, Test paper
The trial-production of fluorescence detection test: based on the colorimetric of namo fluorescence probe of fluorogold nano-cluster and the fluoroscopic examination effect of bovine serum albumin functionalization, it is possible to manufacture experimently into reagent paper, carry out detection application。Filter paper is cut into strip, it is immersed in the namo fluorescence probe aqueous solution of dilution 10～20 times 1～3 hour, after taking-up, filter paper is dried, it is irradiated under uviol lamp, it is observed that strong fluorescence, illustrate filter paper has adsorbed fully BSA-GNPs nano-particle, it is possible to apply as detection instrument reagent paper。Respectively 10-3The TNT of M, paranitrophenol, meta-nitrotoluene, picric acid, 2, the acetone soln of 4-dinitrotoluene (DNT), toluene and Nitrobenzol drops on reagent paper, it is placed under uviol lamp after reagent paper dries and irradiates, find that the reagent paper fluorescence intensity of dropping TNT acetone soln substantially weakens, and the reagent paper of other solution is without significant change, fluorescence is still strong。It is 10 by concentration-3、5×10-4、10-4、5×10-5、10-5、5×10-6、10-6、5×10-7、10-7The TNT solution of M drops on reagent paper respectively, under ultraviolet source, it has been found that dropping 10-3、5×10-4、10-4、5×10-5The filter paper fluorescence intensity of M solution substantially weakens in gradient, TNT solution concentration is more high, and fluorescence is more weak, illustrates that testing result is also with concentration change, consistent with BSA-GNPs namo fluorescence probe solution testing result, thus proving that reagent paper has the Detection results same with BSA-GNPs namo fluorescence probe。
Present invention have the advantage that
1, fluorescent detection probe provided by the invention is highly sensitive, selectivity good, and detection limit is low。
2, large-scale instrument is not needed, by naked eye Colorimetric results or fluorescence spectrum, can recognition detection result。
3, the present invention easily prepares and preserves；10～15 months can be preserved when 4 ° of C。
4, agents useful for same of the present invention and operating process all have no side effect。
5, the inventive method is simply, quickly, easily operate, and can carry out on-the-spot original position and quickly detect。
Accompanying drawing explanation
Accompanying drawing 1, BSA-GNPs namo fluorescence probe the picture of colorimetric detection TNT, from left to right the concentration of TNT is 10-3、5×10-4、10-4、5×10-5、10-5、5×10-6、10-6、5×10-7、10-7M；
The fluorescence picture of accompanying drawing 2, BSA-GNPs namo fluorescence probe detection TNT, from left to right the concentration of TNT is 10-3、5×10-4、10-4、5×10-5、10-5、5×10-6、10-6、5×10-7、10-7M；
The fluorescence picture of the detection paper TNT that accompanying drawing 3, BSA-GNPs namo fluorescence probe are made, from left to right the concentration of TNT is 10-3、5×10-4、10-4、5×10-5、10-5、5×10-6、10-6、5×10-7、10-7M；
The ultraviolet spectrogram of accompanying drawing 4, BSA-GNPs namo fluorescence probe detection TNT；
The fluorescence spectrum figure of accompanying drawing 5, BSA-GNPs namo fluorescence probe detection TNT。
Detailed description of the invention
Embodiment 1:
The synthesis of the fluorogold nano-cluster (BSA-GNanoclusters) of fluorescent probe bovine serum albumin functionalization:
The Nano-Au probe of bovine serum albumin (BSA) functionalization is [XieJP prepared by the method according to bibliographical information, ZhengYG, YingJY, Protein-DirectedSynthesisofHighlyFluorescentGoldNanoclus ters [J] .J.Am.Chem.Soc.2009,131 (3): 888 889.]。Concrete synthetic method is as follows: gold chloride is made into the stock solution of 10mM, under the water bath condition of 30～40 ° of C, it is that 1.5～3:1 is made into aqueous solution by the mass ratio of bovine serum albumin Yu two kinds of reactants of gold chloride, two kinds of solution mix under magnetic stirring, after 2～5 minutes, the NaOH solution adding the 1M accounting for mixed liquor volume 5% regulates the pH value of mixed solution, then under 30～40 ° of C water bath condition of constant temperature continuously stirred 10～24 hours。The color of solution is become dark-brown by the glassy yellow of original chlorauric acid solution。Fluorogold nano-cluster solution presents strong red emission under ultraviolet source, excites under the wavelength of 250～450nm, and fluorogold nano-cluster has strong emission spectrum at 630～660nm。
Embodiment 1:
The BSA-GNPs solution prepared has very strong fluorescent emission, and in order to make the sensitivity in detection process improve, original solution is diluted 10 times, and the BSA-GNPs solution after the dilution obtained is taken as nanometer detection probe。In the experiment of inspection BSA-GNPs nano-probe sensitivity, taking concentration respectively is 10-3、5×10-4、10-4、5×10-5、10-5、5×10-6、10-6、5×10-7、10-7The TNT acetone soln of M is as testing sample, and sample to be measured and BSA-GNPs fluorescent detection probe are the amount hybrid reaction of 1:1 by volume, it can be observed that 10-3、5×10-4、10-4、5×10-5The color generation significant change of the TNT of M concentration, color is from the original colourless redness that becomes, and the concentration along with TNT of the color of solution raises and deepens；Meanwhile, under the irradiation of uviol lamp, fluorescence intensity dies down successively。Pass through fluorescence spectrum, it is possible to verify the sensitivity of probe further。Therefore, BSA-GNPs can be used for detecting TNT as colorimetric and fluorescent dual probe。
Embodiment 2:
Filter paper is cut into strip, is immersed in the BSA-GNPs aqueous solution of dilution 10 times 2 hours, after taking-up, filter paper is dried, it is irradiated under uviol lamp, it is observed that strong fluorescence, illustrate filter paper has adsorbed fully BSA-GNPs nano-particle, it is possible to apply as detection instrument reagent paper。Respectively 10-3The TNT of M, paranitrophenol, meta-nitrotoluene, picric acid, 2, the acetone soln of 4-dinitrotoluene (DNT), toluene and Nitrobenzol drops on reagent paper, it is placed under the ultraviolet source of 254nm after reagent paper dries and observes, find that the reagent paper fluorescence intensity of dropping TNT acetone soln substantially weakens, and the reagent paper of other solution is without significant change, fluorescence is still strong。It is 10 by concentration-3、5×10-4、10-4、5×10-5、10-5、5×10-6、10-6、5×10-7、10-7The TNT solution of M drops on reagent paper respectively, under ultraviolet source, it has been found that dropping 10-3、5×10-4、10-4、5×10-5The filter paper fluorescence intensity of M solution substantially weakens in gradient, and TNT solution concentration is more high, and fluorescence is more weak, illustrates that testing result presents rule change with concentration, consistent with BSA-GNPs namo fluorescence probe solution testing result。
Claims (1)
1. the method that the fluorogold nano-cluster of bovine serum albumin functionalization detects TNT as Test paper, it is characterized in that: filter paper is cut into strip, it is immersed in the fluorogold nano-cluster BSA-GNPs aqueous solution of the bovine serum albumin functionalization diluting 10 times 2 hours, after taking-up, filter paper is dried, respectively 10-3The TNT of M, paranitrophenol, meta-nitrotoluene, picric acid, 2, the acetone soln of 4-dinitrotoluene (DNT), toluene and Nitrobenzol drops on reagent paper, it is placed under the ultraviolet source of 254nm after reagent paper dries and observes, find that the reagent paper fluorescence intensity of dropping TNT acetone soln substantially weakens, and the reagent paper of other solution is without significant change, fluorescence is still strong, the TNT solution of variable concentrations is dropped on reagent paper respectively, under ultraviolet source, find that the filter paper fluorescence intensity of dropping solution substantially weakens in gradient, TNT solution concentration is more high, and fluorescence is more weak。
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| CN109540852B (en) * | 2017-09-22 | 2022-03-25 | 清水湾生物材料(深圳)有限公司 | Fluorescent detection test paper, preparation method and application thereof, and inferior grease identification method |
| CN109839500A (en) * | 2017-11-24 | 2019-06-04 | 中国农业大学 | A kind of label-free fluorescence enzyme-linked immune analytic method based on fluorescence gold nanoclusters |
| CN111157506B (en) * | 2020-01-17 | 2022-10-21 | 天津师范大学 | Method for detecting thioglycollic acid by using integrated fluorescent test paper |
| CN113136205A (en) * | 2021-04-12 | 2021-07-20 | 广东石油化工学院 | Fluorescent carbon quantum dot, preparation method and application thereof in detecting superoxide anion |
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