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A "fluorescence-stable isotope" dual-mode detection method for p-trinitrotoluene

A stable isotope, trinitrotoluene technology, used in fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of difficult long-term stable and accurate quantitative analysis, limited photobleaching effect and matrix effect, etc., to achieve high-sensitivity detection. , long-term stability and high sensitivity detection, the effect of small matrix effect

Active Publication Date: 2021-07-16
SICHUAN UNIV
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  • Description
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  • Application Information

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Problems solved by technology

However, the traditional fluorescence spectroscopy is limited by the photobleaching effect and matrix effect, and it is difficult to achieve long-term stable and accurate quantitative analysis. Therefore, in the field of TNT analysis, there is still a need for a fast and convenient on-site detection of TNT, which can also ensure accuracy and stability. Highly Sensitive Quantitative Method

Method used

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  • A "fluorescence-stable isotope" dual-mode detection method for p-trinitrotoluene
  • A "fluorescence-stable isotope" dual-mode detection method for p-trinitrotoluene
  • A "fluorescence-stable isotope" dual-mode detection method for p-trinitrotoluene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1, whether to form the comparative experiment of terminal group protection to TNT detection

[0059] In order to confirm that the formation of end group protection at the end of TNT and AptTNT-T30 aptamer is the key to the synthesis of PolyT CuNPs, the AptTNT-T30 probe was used as an example to respond to TNT, and compared the formation of end group protection with or without the presence of ExoⅠ enzyme on the synthesis of PolyT CuNPs. Effect of Fluorescent Properties of Poly-CuNPs. In the experiment, add TNT sample 200nM, ExoI 40U, excitation wavelength 340nm, emission wavelength 660nm. Depend on figure 1 It can be seen that when there is no TNT sample, no end group protection is formed. After adding ExoⅠ enzyme, the digestion reaction with AptTNT-T30 occurs, and the fluorescence signal response is significantly weakened, which is close to blank, and PolyT CuNPs cannot be formed. However, adding inactivated ExoⅠ When the enzyme was used, the fluorescence r...

Embodiment 2

[0060] Embodiment 2, exploring the specificity detection of TNT by the analytical method of the present invention

[0061] This example uses the "fluorescence-stable isotope" dual-mode p-trinitrotoluene detection method to detect various interfering substances (other common explosives, metal ions, amino acids and biological enzymes), and explores the analysis method of the present invention Specific detection ability for TNT.

[0062] 1. Detection of TNT and other interfering substances

[0063] Using the specific steps above, TNT and various interfering substances were detected in the fluorescence detection mode and the stable isotope detection mode, respectively.

[0064] Among them, in the fluorescence detection mode, first react 10 μL, 10 μM AptTNT-T30 substrate and 10 μL test substance in a PCR instrument. Among them, the concentration of TNT to be tested is 20nM, other interfering substances are 1mM, and the reaction conditions are 25°C, 90min. Then add 25 μL Nebuffer...

Embodiment 3

[0068] Embodiment 3, exploring the sensitivity detection of the analytical method of the present invention to TNT

[0069] This example explores the sensitivity and linearity of TNT detection in the fluorescence detection mode and stable isotope detection mode.

[0070] 1. The sensitivity detection of analytical method of the present invention to TNT

[0071] In the fluorescence detection mode, first react 10 μL, 10 μM AptTNT-T30 substrate and 10 μL TNT of different concentrations in a PCR instrument. Among them, the concentration of TNT to be tested is 20nM, other interfering substances are 1mM, and the reaction conditions are 25°C, 90min. Then add 25 μL Nebuffer 3 buffer solution and 40U Exo I for enzymatic hydrolysis reaction. The PCR reaction conditions were 37° C., 120 min. After the enzymatic hydrolysis reaction, the temperature was raised to 85°C for 15 minutes to inactivate the enzyme. Finally, 10 μL of 20 mM ascorbic acid solution and 35 mL of 1 mM copper sulfate ...

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Abstract

The purpose of the present invention is to provide a "fluorescence-stable isotope" dual-mode detection method for p-trinitrotoluene (TNT). The assay consists of a polythymidine‑nucleic acid aptamer probe (AptTNT‑T30) and dual-modality detection with fluorescence detection and stable isotope detection. The principle of the present invention is: the TNT nucleic acid aptamer (AptTNT) sequence can be specifically combined with the target TNT molecule for terminal protection, avoiding exonuclease cutting, and the polythymine sequence (T30) at the 5' end can be used as a template Synthesis of copper nanoparticles. The synthesized copper nanoparticles can be detected in dual modes: in the fluorescence detection mode, rapid fluorescence detection can be performed; after the copper nanoparticles are digested in nitric acid, in the stable isotope detection mode, the inductively coupled plasma mass spectrometer can be used for highly sensitive and stable quantitative analysis , which can respectively satisfy the on-site rapid detection and highly sensitive and accurate detection of TNT.

Description

technical field [0001] The invention belongs to the detection field of analytical chemistry, relates to the field of Fluorimetry and stable isotope (Metal stable isotope) sensing of explosives, and particularly relates to an in-situ detection method based on polythymine deoxyribonucleic acid (Poly-T ssDNA) as a template Synthetic Copper Nanoparticles (CuNPs) Fluorescence-Stable Isotope Bimodal Paratrinitrotoluene (TNT) Detection Method. Background technique [0002] Trinitrotoluene (TNT), as one of the most typical chemical explosives, has been widely used in military, mining, anti-terrorism and civilian fields. At the same time, TNT is a highly toxic substance, which poses a serious toxicological threat to animals, plants and humans. The serious pollution of TNT to soil and water caused by widespread application threatens the survival of ecosystems. In addition, the increasingly severe global security situation and anti-terrorism requirements are becoming more and more im...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 吕弋胡建煜刘睿
Owner SICHUAN UNIV
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