CN103424388A - TNT colorimetric and fluorescent detection probe and application method thereof - Google Patents
TNT colorimetric and fluorescent detection probe and application method thereof Download PDFInfo
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- CN103424388A CN103424388A CN2013102527039A CN201310252703A CN103424388A CN 103424388 A CN103424388 A CN 103424388A CN 2013102527039 A CN2013102527039 A CN 2013102527039A CN 201310252703 A CN201310252703 A CN 201310252703A CN 103424388 A CN103424388 A CN 103424388A
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Abstract
The invention belongs to the technical field of nano detection and particularly relates to a 2, 4, 6-trinitrotoluene (TNT) colorimetric and fluorescent difunctional detection probe and an application method thereof. A gold nano-cluster for Bovine serum albumin (BSA) functionalization is provided as a colorimetric and dual-detection probe, a diluted BSA-GNPs (gold nanoparticles) solution serves as a nano detection probe and performs hybrid reaction with a sample to be tested at the volume ratio of 1:1, and then the TNT color change is observed. Also, a piece of filter paper can be cut into strips and soaked in the BSA-GNPs water solution which is diluted by 10 times, and then the strips are taken out and dried to serve as detection test paper. The strips can also serve as a colorimetric detection probe, and the existence of TNT can be detected through the colorimetric method under the ordinary sunlight condition. In-situ quick detection can be realized, the detection limit is low, the TNT colorimetric and fluorescent detection probe can be applied under more conditions and within a larger range, the operation is simple, the manufacturing cost is low, and the reagents and the operation process have no toxic and adverse effects.
Description
Technical field
The invention belongs to the nanometer detection technical field, be specifically related to colorimetric and fluorescent dual-function detector probe and the application process thereof of a kind of TNT (TNT).
Background technology
TNT (Trinitrotoluene, TNT, 2,4, be 6-trinitromethylbenzene) a kind of faint yellow aromatic hydrocarbons crystal, molten point is 354 K, it is one of Explosive ingredients commonly used with explosivity.The trinitro-toluene of refining is very stable.Different from monobel, it is all insensitive for friction, vibrations etc.But it and alkali reaction kickback, generate unsettled compound.These compounds are all very responsive to heat and shock.The toxicity of TNT is moderate toxicity, can invade through skin, respiratory tract, alimentary canal, and main harm is slow poisoning, and people's long term exposure can increase and suffer from anemia and the abnormal chance of liver function in trinitro-toluene; The animal of having injected or sucked trinitro-toluene also discovery can affect blood and liver, spleen and sends out greatly and other You Guan immune bad influences; Also proved on evidence that TNT has bad influence to the male sex's reproductive function, and TNT also be listed in a kind of may carcinogenic substance.Feed TNT can make the urine blackening.Some military test base is polluted by TNT.The sewage that munitions produce can pollute the day water and underground water.These contaminated water meeting pinkiness, this is because water is polluted by TNT.These pollutants are called as pink water, clear up that it is very difficult and expensive.
Therefore, in recent years, in the exploration of the detection to the ultratrace nitroaromatic and relevant sensor array, caused that society research mechanism pays close attention to and fruitful exploration widely.The laboratory of specific nitryl aromatic family's explosive and vapor signal thereof is surveyed and is carried out widely by the method for gas chromatograph-mass spectrometer, ion mobility spectrometry and neutron activity analysis etc.These traditional analysis technology can meet the basic demand in analysis, as selectivity, reliability, accuracy and repeatability, but these detection methods are expensive, consuming time and loaded down with trivial details heaviness, because in detecting, sample must be to break away from Test Field to be sent to laboratory and to go to analyze, can not accomplish the detection of real-time on-site.In sum, be necessary to find a kind of can be fast and detect easily the method for TNT.
For above problem, we have researched and developed a kind of gold nanoclusters of functionalization that utilizes and have detected the method for TNT with the double function probe that is colorimetric and fluoroscopic examination.The invention provides a kind ofly for detection of the colorimetric of TNT and the double function probe of fluoroscopic examination, and provide that it is simple to operate, the method for Quantitative detection TNT.
Summary of the invention
The colorimetric and the fluorescent dual-function detector probe that the purpose of this invention is to provide a kind of explosive detection TNT.
Another object of the present invention is to provide the application process of the detector probe of a kind of explosive detection TNT.
The fluorogold nano-cluster that fluorescent detection probe for mercury ions of the present invention is bovine serum albumin(BSA) (BSA) functionalization.
The bovine serum albumin(BSA) molecular surface is rich in each seed amino acid, contain amino and carboxylic group with the nanogold particle rich surface of bovine serum albumin(BSA) functionalization, the nitryl aromatic explosive TNT that has the aromatic rings of electron deficient is the acceptor of an electronics, the fluorescent material surface so just demonstrated electron rich has very high affinity, this photoluminescence shifts the direct cancellation of two compound mechanism at electron accepter with between body by electronics, this cancellation mainly depends on the electronic capability of accepting of nitryl aromatic compound, therefore, pass through fluorescence detection method, can detect the TNT of the trace contained in sample.
One, the fluorogold nano-cluster (BSA-GNanoclusters) of fluorescence probe bovine serum albumin(BSA) functionalization is synthetic:
The Nano-Au probe of bovine serum albumin(BSA) (BSA) functionalization is [Xie J P prepared by the method according to bibliographical information, Zheng Y G, Ying J Y, Protein-Directed Synthesis of Highly Fluorescent Gold Nanoclusters [J] .J. Am. Chem. Soc. 2009,131 (3): 888 – 889.].Concrete synthetic method is as follows: the stock solution that gold chloride is made into to 10 mM, under the water bath condition of 30～40 ° of C, mass ratio by bovine serum albumin(BSA) and two kinds of reactants of gold chloride is that 1.5～3:1 is made into aqueous solution, two kinds of solution mix under magnetic agitation, after 2～5 minutes, the pH value that adds the NaOH solution adjusting mixed solution of the 1M that accounts for mixed liquor volume 5% then continues to stir 10～24 hours under 30～40 ° of C water bath condition of constant temperature.The color of solution has become dark-brown by the glassy yellow of original chlorauric acid solution.Fluorogold nano-cluster solution presents strong red emission under ultraviolet source, under the wavelength of 250～450 nm, excites, and the fluorogold nano-cluster has strong emission spectrum at 630～660 nm.
Two, detect TNT
Testing process: the fluorogold nano-cluster of the bovine serum albumin(BSA) functionalization prepared (hereinafter to be referred as the fluorogold nano-cluster) solution has very strong fluorescent emission, for the sensitivity in testing process is improved, original solution is diluted 10～20 times, and the fluorogold nano-cluster solution after the dilution obtained is used as the nanometer detection probe.The gold nanoclusters detector probe of bovine serum albumin(BSA) functionalization not only can come by the variation of fluorescence indicating target to detect the existence of thing TNT, can also be as colorimetric detection probes, do not need uviol lamp to excite, under common sunshine condition, can go out by colorimetric determination the existence of TNT.In the process of fluorogold nano-cluster detector probe explosive detection, sample to be measured mixes for the amount of 1:1 by volume with fluorogold nano-cluster fluorescent detection probe, the solution example of the explosive of 100 μ L adds in 100 μ L nanometer detection probes, all sensitivity experiments and selectivity experiment are all carried out in above ratio, and concrete experimentation is as follows:
1, detection sensitivity
The fluorogold nano-cluster detector probe of using the bovine serum albumin(BSA) functionalization during as nanometer detection probe in detecting TNT, has very high sensitivity.
In the experiment of check fluorogold nano-cluster detector probe sensitivity, getting respectively concentration 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 100 μ L of M react with the fluorogold nano-cluster aqueous solution of 10 times of the dilutions of equivalent, can observe 10
-3, 5 * 10
-4, 10
-4, 5 * 10
-5The color generation significant change of the TNT of M concentration, color is by the original colourless redness that becomes, and the concentration along with TNT of the color of solution raises and deepens; Simultaneously, under the irradiation of uviol lamp, with the rising fluorescence intensity of TNT concentration, die down successively.By ultraviolet spectrum and fluorescence spectrum, can further verify the sensitivity (accompanying drawing 4, accompanying drawing 5) of probe.Therefore, the nanometer detection probe can be used as colorimetric and fluorescent dual-function probe for detection of TNT.
2, selectivity
In the fluorogold nano-cluster namo fluorescence probe detection TNT of check bovine serum albumin(BSA) functionalization optionally tests, by p-nitrophenol, meta-nitrotoluene, picric acid, 2, the explosive of 4-dinitrotoluene (DNT), toluene and nitrobenzene and TNT structural similarity, as detected object, is investigated the selectivity of detector probe.Getting respectively concentration is 10
-3The TNT of M, p-nitrophenol, meta-nitrotoluene, picric acid, 2, the fluorogold nano-cluster solution reaction that the acetone soln 100 μ L of 4-dinitrotoluene (DNT) and 100 μ L dilution are 10～20 times, can see that the fluorogold nano-cluster solution colour that adds TNT changes, become redness, and other solution colours except p-nitrophenol becomes yellow without significant change.While irradiating under ultraviolet source, the fluorescence intensity of TNT dies down, and the fluorescence intensity of other solution does not have significant change, therefore, can judge namo fluorescence probe the detection of TNT is had to selectivity preferably.
3, Test paper
The trial-production of fluorescence detection test: the colorimetric of the namo fluorescence probe of the fluorogold nano-cluster based on the bovine serum albumin(BSA) functionalization and fluoroscopic examination effect, can manufacture experimently into test paper, detect application.Filter paper is cut into to strip, be immersed in the dilution namo fluorescence probe aqueous solution of 10～20 times 1～3 hour, after taking-up, filter paper is dried, it is irradiated under uviol lamp, can observe strong fluorescence, illustrate in filter paper and adsorbed fully the BSA-GNPs nano particle, can be used as testing tool-test paper and apply.Respectively 10
-3The TNT of M, p-nitrophenol, meta-nitrotoluene, picric acid, 2, the acetone soln of 4-dinitrotoluene (DNT), toluene and nitrobenzene drops on test paper, be placed on after test paper dries under uviol lamp and irradiate, find that the test paper fluorescence intensity that drips the TNT acetone soln obviously weakens, and the test paper of other solution is without significant change, fluorescence is still strong.By concentration, be 10
-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 in respectively on test paper, under ultraviolet source, finds to drip 10
-3, 5 * 10
-4, 10
-4, 5 * 10
-5The filter paper fluorescence intensity of M solution obviously weakens in gradient, the TNT solution concentration is higher, 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 the proof test paper has the detection effect same with the BSA-GNPs namo fluorescence probe.
The present invention has following advantage:
1, fluorescent detection probe provided by the invention is highly sensitive, selectivity good, and detectability is low.
2, do not need large-scale instrument, by bore hole, observe colorimetric result or fluorescence spectrum, can the recognition detection result.
3, the present invention easily prepares and preserves; Under 4 ° of C conditions, can preserve 10～15 months.
4, agents useful for same of the present invention and operating process all have no side effect.
5, the inventive method is simple, quick, easy to operate, can carry out on-the-spot original position fast detecting.
The accompanying drawing explanation
The picture of the colorimetric detection TNT of accompanying drawing 1, BSA-GNPs namo fluorescence probe, 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;
Accompanying drawing 2, BSA-GNPs namo fluorescence probe detect the fluorescence picture of TNT, and 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;
Accompanying drawing 4, BSA-GNPs namo fluorescence probe detect the ultraviolet spectrogram of TNT;
Accompanying drawing 5, BSA-GNPs namo fluorescence probe detect the fluorescence spectrum figure of TNT.
Embodiment
Embodiment 1:
Synthesizing of the fluorogold nano-cluster (BSA-GNanoclusters) of fluorescence probe bovine serum albumin(BSA) functionalization:
The Nano-Au probe of bovine serum albumin(BSA) (BSA) functionalization is [Xie J P prepared by the method according to bibliographical information, Zheng Y G, Ying J Y, Protein-Directed Synthesis of Highly Fluorescent Gold Nanoclusters [J] .J. Am. Chem. Soc. 2009,131 (3): 888 – 889.].Concrete synthetic method is as follows: the stock solution that gold chloride is made into to 10 mM, under the water bath condition of 30～40 ° of C, mass ratio by bovine serum albumin(BSA) and two kinds of reactants of gold chloride is that 1.5～3:1 is made into aqueous solution, two kinds of solution mix under magnetic agitation, after 2～5 minutes, the pH value that adds the NaOH solution adjusting mixed solution of the 1M that accounts for mixed liquor volume 5% then continues to stir 10～24 hours under 30～40 ° of C water bath condition of constant temperature.The color of solution has become dark-brown by the glassy yellow of original chlorauric acid solution.Fluorogold nano-cluster solution presents strong red emission under ultraviolet source, under the wavelength of 250～450 nm, excites, and the fluorogold nano-cluster has strong emission spectrum at 630～660 nm.
Embodiment 1:
The BSA-GNPs solution prepared has very strong fluorescent emission, in order to make the sensitivity in testing process, improves, and original solution is diluted 10 times, and the BSA-GNPs solution after the dilution obtained is used as the nanometer detection probe.In the experiment of check BSA-GNPs nano-probe sensitivity, getting respectively concentration 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, can observe 10
-3, 5 * 10
-4, 10
-4, 5 * 10
-5The color generation significant change of the TNT of M concentration, color is by the original colourless redness that becomes, and the concentration along with TNT of the color of solution raises and deepens; Simultaneously, under the irradiation of uviol lamp, fluorescence intensity dies down successively.By fluorescence spectrum, can further verify the sensitivity of probe.Therefore, BSA-GNPs can be used as colorimetric and fluorescent dual probe for detection of TNT.
Embodiment 2:
Filter paper is cut into to strip, is immersed in the dilution BSA-GNPs aqueous solution of 10 times 2 hours, after taking-up, filter paper is dried, it is irradiated under uviol lamp, can observe strong fluorescence, illustrate in filter paper and adsorbed fully the BSA-GNPs nano particle, can be used as testing tool-test paper and apply.Respectively 10
-3The TNT of M, p-nitrophenol, meta-nitrotoluene, picric acid, 2, the acetone soln of 4-dinitrotoluene (DNT), toluene and nitrobenzene drops on test paper, be placed on after test paper dries under the ultraviolet source of 254 nm and observe, find that the test paper fluorescence intensity that drips the TNT acetone soln obviously weakens, and the test paper of other solution is without significant change, fluorescence is still strong.By concentration, be 10
-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 in respectively on test paper, under ultraviolet source, finds to drip 10
-3, 5 * 10
-4, 10
-4, 5 * 10
-5The filter paper fluorescence intensity of M solution obviously weakens in gradient, and the TNT solution concentration is higher, and fluorescence is more weak, illustrates that testing result presents rule with concentration and changes, consistent with BSA-GNPs namo fluorescence probe solution testing result.
Claims (3)
- The fluorogold nano-cluster of bovine serum albumin(BSA) functionalization as fluorescence probe in the application detected aspect TNT.
- 2. the fluorogold nano-cluster of bovine serum albumin(BSA) functionalization detects the method for TNT as fluorescence probe, it is characterized in that: the BSA-GNPs original solution is diluted to 10 times, as the nanometer detection probe, get respectively the TNT acetone soln of variable concentrations as testing sample, sample to be measured and BSA-GNPs fluorescent detection probe are the amount hybrid reaction of 1:1 by volume, observe 10 -3, 5 * 10 -4, 10 -4, 5 * 10 -5The color generation significant change of the TNT of M concentration, color is by the original colourless redness that becomes, and the concentration along with TNT of the color of solution raises and deepens; Simultaneously, under the irradiation of uviol lamp, fluorescence intensity dies down successively, by fluorescence spectrum, can further verify the sensitivity of probe.
- 3. the fluorogold nano-cluster of bovine serum albumin(BSA) functionalization detects the method for TNT as Test paper, it is characterized in that: filter paper is cut into to strip, is immersed in the BSA-GNPs aqueous solution of 10 times of dilutions 2 hours, after taking-up, filter paper is dried, respectively 10 -3The TNT of M, p-nitrophenol, meta-nitrotoluene, picric acid, 2, the acetone soln of 4-dinitrotoluene (DNT), toluene and nitrobenzene drops on test paper, be placed on after test paper dries under the ultraviolet source of 254 nm and observe, find that the test paper fluorescence intensity that drips the TNT acetone soln obviously weakens, and the test paper of other solution is without significant change, fluorescence is still strong, by the TNT solution of variable concentrations, drop in respectively on test paper, under ultraviolet source, find that the filter paper fluorescence intensity that drips solution obviously weakens in gradient, the TNT solution concentration is higher, and fluorescence is more weak.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109540852A (en) * | 2017-09-22 | 2019-03-29 | 惠州清水湾生物材料有限公司 | Fluorescence detection test and its preparation method and application, grease identification method inferior |
| 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 |
| CN111157506A (en) * | 2020-01-17 | 2020-05-15 | 天津师范大学 | Integrated fluorescent test paper for detecting thioglycollic acid in real time and application |
| 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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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109540852A (en) * | 2017-09-22 | 2019-03-29 | 惠州清水湾生物材料有限公司 | Fluorescence detection test and its preparation method and application, grease identification method inferior |
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| CN111157506A (en) * | 2020-01-17 | 2020-05-15 | 天津师范大学 | Integrated fluorescent test paper for detecting thioglycollic acid in real time and application |
| 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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