Dual-target fluorescent probes for detecting nerve agents and cyanide and applications thereof

Abstract

The application discloses a double-target fluorescent probe for detecting nerve agents and cyanides, and a structural general formula is shown as formula I: the double-target fluorescent probe can detect nerve agents and cyanides simultaneously, and the reaction rate is fast. After the double-target fluorescent probe reacts with nerve agents, the fluorescence can be significantly quenched; after the double-target fluorescent probe reacts with cyanides, the fluorescence can be caused to blue shift.

Description

Technical Field

[0001] This invention belongs to the field of biological detection technology, specifically, it relates to a dual-target fluorescent probe for detecting nerve agents and cyanide and its application. Background Technology

[0002] Nerve agents are a class of highly toxic organophosphate compounds, including tabun, sarin, soman, and VX. Due to their high toxicity, diethyl chlorophosphate (DCP) is generally used in experiments as a substitute for nerve agents. Nerve agents primarily work by inhibiting the activity of acetylcholinesterase, leading to its accumulation in the synaptic cleft, further causing excessive excitation of the nervous system, and ultimately resulting in death. Systemic agents are also known as cyanides. Billions of tons of cyanide are used in industrial production worldwide each year; therefore, rapid detection of nerve agents and cyanides is of great importance for maintaining public safety and protecting public health.

[0003] Currently, traditional analytical methods are still used in actual testing operations, mainly including mass spectrometry, titration, chromatography, and electrochemical methods. These methods require large-scale instruments, which are not portable and are difficult to meet the needs of routine anti-toxic testing, such as in public places like train stations and airports where people gather, to achieve real-time and rapid anti-toxic testing. Furthermore, their detection limits are limited, their accuracy is limited, and their testing time is relatively long, making them difficult to promote in practice.

[0004] In recent years, the rapid development of small molecule fluorescent probe technology has made it possible to quickly and sensitively detect nerve agents and cyanide. This technology has attracted widespread interest from researchers and has been widely used in medicine, chemistry, and biological sciences, gradually becoming an indispensable research tool in medical diagnostics, life sciences, and environmental sciences. However, currently reported small molecule fluorescent probes can only detect nerve agents or systemic poisons, and cannot simultaneously detect both types of fast-acting agents. Summary of the Invention

[0005] The purpose of this invention is to provide a dual-target fluorescent probe for detecting nerve agents and cyanides.

[0006] Another object of the present invention is to provide the application of the dual-target fluorescent probe in the preparation of fluorescent detection reagents for nerve agents or cyanide.

[0007] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0008] In a first aspect, the present invention provides a dual-target fluorescent probe for detecting nerve agents and cyanide, having the general structural formula shown in Formula I:

[0009]

[0010] R1 is selected from hydrogen and C1-C5 alkyl groups;

[0011] R2 is selected from hydrogen, C1-C5 alkyl groups,

[0012] R3 is selected from hydrogen and C1-C5 alkyl groups;

[0013] Alternatively, R2 can form a six-membered ring with R1, C, and N; and R2 can also form a six-membered ring with R3, C, and N.

[0014] Preferably, in the dual-target fluorescent probe,

[0015] R1 is selected from hydrogen, methyl, and ethyl.

[0016] R2 is selected from hydrogen, methyl, ethyl,

[0017] R3 is selected from hydrogen, methyl, and ethyl;

[0018] Alternatively, the structure formed by R2 with R1 and R3 may have the following groups: The dashed line indicates the connection point.

[0019] Most preferably, the structure of the dual-target fluorescent probe for detecting nerve agents and cyanide is selected from one of the following structures:

[0020]

[0021] A second aspect of the present invention provides the application of the dual-target fluorescent probe in the preparation of fluorescent detection reagents for nerve agents and / or fluorescent detection reagents for cyanides.

[0022] The dual-target fluorescent probe of this invention can not only detect nerve agents and cyanide in biological and environmental samples, but also be used to detect the concentration of nerve agents and cyanide in living cells.

[0023] The application of the dual-target fluorescent probe as a reagent for the fluorescence detection of nerve agents includes the following steps:

[0024] The dual-target fluorescent probe solution was added to the sample solution to achieve a final concentration of 20 μM. After incubation for 1 min, the sample was excited at 365 nm, and the fluorescence intensity of the solution was recorded at 582 nm or 596 nm. The dual-target fluorescent probe solution was used as a blank control. If the sample contained nerve agents, the fluorescence intensity at 582 nm or 596 nm was lower than that of the blank control. If the sample did not contain nerve agents, the fluorescence intensity at 582 nm or 596 nm remained unchanged compared with the blank control.

[0025] The application of the dual-target fluorescent probe as a cyanide fluorescence detection reagent includes the following steps:

[0026] The dual-target fluorescent probe solution was added to the sample solution to make the final concentration of the dual-target fluorescent probe 20 μM. After incubation for 1 min, the solution was excited at 365 nm, and the fluorescence intensity at 582 nm and 405 nm was recorded, or the fluorescence intensity at 596 nm and 403 nm was recorded.

[0027] Using the dual-target fluorescent probe solution as a blank control, if the sample contains cyanide, the fluorescence intensity at 582 nm is weaker than that of the blank control, while the fluorescence intensity at 405 nm is stronger; if the sample does not contain cyanide, the fluorescence intensity at 582 nm remains unchanged compared to the blank control, and the fluorescence intensity at 405 nm also remains unchanged compared to the blank control.

[0028] Alternatively, a dual-target fluorescent probe solution can be used as a blank control. If the sample contains cyanide, the fluorescence intensity at 596 nm will be weaker than that of the blank control, while the fluorescence intensity at 403 nm will be stronger. If the sample does not contain cyanide, the fluorescence intensity at 596 nm will remain unchanged compared to the blank control, and the fluorescence intensity at 403 nm will also remain unchanged compared to the blank control.

[0029] The samples to be tested are selected from organic solvents containing nerve agents (such as acetonitrile, dimethyl sulfoxide, methanol, etc.), soil containing nerve agents, or living biological cells containing nerve agents.

[0030] The nerve agent is selected from sarin, soman, tabun, diethyl chlorophosphate, diethyl cyanophosphate, etc.

[0031] The samples to be tested are selected from solvents containing cyanide (such as methanol and water), soil containing cyanide, or living biological cells containing cyanide.

[0032] The cyanide is selected from KCN.

[0033] A third aspect of the present invention provides the application of the dual-target fluorescent probe in the preparation of a fluorescent detection element for nerve agents or a fluorescent detection element for cyanide.

[0034] The nerve agent fluorescence detection element or cyanide fluorescence detection element is selected from fluorescent test strips or kits, wherein the fluorescent test strips are used for qualitative detection and the kits are used for quantitative detection.

[0035] By adopting the above technical solution, the present invention has the following advantages and beneficial effects:

[0036] The dual-target fluorescent probe of this invention has high sensitivity, with a detection limit of 5.5 μM for nerve agents and 9.6 nM for cyanide.

[0037] The dual-target fluorescent probe of this invention has a fast reaction rate, and can react with nerve agents within 10 seconds and with cyanide within 1 minute.

[0038] The dual-target fluorescent probe of this invention has good selectivity, reacting only with nerve agents and cyanide, and not with other common chemical agents such as mustard gas and Lewis agents.

[0039] The dual-target fluorescent probe of this invention can simultaneously detect nerve agents and cyanide, and has a fast reaction rate. The dual-target fluorescent probe of this invention can significantly quench fluorescence upon reaction with nerve agents; and it can induce a blue shift in fluorescence upon reaction with cyanide. Attached Figure Description

[0040] Figure 1 This is a schematic diagram of the proton NMR spectrum of fluorescent probe I-1.

[0041] Figure 2 This is a schematic diagram of the high-resolution mass spectrum of fluorescent probe I-1.

[0042] Figure 3 This is a schematic diagram of the proton NMR spectrum of fluorescent probe I-2.

[0043] Figure 4 This is a schematic diagram of the high-resolution mass spectrum of fluorescent probe I-2.

[0044] Figure 5 This is a schematic diagram showing the fluorescence changes of fluorescent probe I-1 before and after reacting with different concentrations of nerve agents.

[0045] Figure 6 This is a schematic diagram showing the fluorescence changes of fluorescent probe I-1 before and after reacting with different concentrations of cyanide.

[0046] Figure 7This is a schematic diagram showing the fluorescence changes of fluorescent probe I-2 before and after reacting with different concentrations of nerve agents.

[0047] Figure 8 This is a schematic diagram showing the fluorescence changes of fluorescent probe I-2 before and after reacting with different concentrations of cyanide.

[0048] Figure 9 This is a schematic diagram showing the fluorescence changes of fluorescent probes I-1 and I-2 at different times after reacting with nerve agents.

[0049] Figure 10 This is a schematic diagram showing the fluorescence changes of fluorescent probes I-1 and I-2 at different times after reacting with cyanide.

[0050] Figure 11 This is a schematic diagram illustrating the selectivity of fluorescent probe I-1 for nerve agents.

[0051] Figure 12 This is a schematic diagram showing the selectivity of fluorescent probe I-1 for cyanide.

[0052] Figure 13 This is a schematic diagram of the detection of nerve agents and cyanide by fluorescent probes I-1 and I-2.

[0053] Figure 14 This is a schematic diagram of fluorescence imaging images of live cells incubated with fluorescent probe I-1 reacting with different concentrations of nerve agents or cyanide. Detailed Implementation

[0054] To more clearly illustrate the present invention, the following description, in conjunction with preferred embodiments, further clarifies the invention. Those skilled in the art should understand that the specific descriptions below are illustrative rather than restrictive, and should not be construed as limiting the scope of protection of the present invention.

[0055] Example 1

[0056] Synthesis of fluorescent probe I-1

[0057]

[0058] Benzimidazole-2-acetonitrile (compound C2) (0.27 g, 1.4 mmol) and pyridine (0.05 mL, 0.7 mmol) were added to 50 mL of methanol solution and stirred at room temperature for 30 min. Then, compound C1 (0.29 g, 1.4 mmol) was added and stirred at room temperature for 6 h. The mixture was filtered, washed three times with methanol, and dried to obtain compound C3.

[0059] Compound C3 (0.17 g, 0.5 mmol) and malononitrile (0.03 g, 0.5 mmol) were added to 2-methoxyethanol (25 mL), and the mixture was heated under reflux for 10 h. The reaction solution was cooled to 5 °C, the precipitate was collected, washed with ethanol, and separated to obtain compound I-1.

[0060] Nuclear magnetic resonance and mass spectrometry, such as Figure 1 and Figure 2 As shown, Figure 1 This is a schematic diagram of the proton NMR spectrum of fluorescent probe I-1. Figure 2 This is a schematic diagram of the high-resolution mass spectrum of fluorescent probe I-1. 1 H NMR (400MHz, CDCl3) δ8.74-8.76(m,1H),8.44-8.47(m,2H),7.80-7.82(m,1H),7.41-7.44(m,3H),6.84-6.86(m,2H),3.51(s,4H),1.73(s,6H). ESI-MS m / z:ESI-HRMS(m / z):[M+H] + Calculated for I-1(C 24 H 20 N5O,M + ):394.1668,found394.1670.

[0061] Example 2

[0062] Synthesis of fluorescent probe I-2

[0063]

[0064] Benzimidazole-2-acetonitrile (compound C2) (0.27 g, 1.4 mmol) and pyridine (0.05 mL, 0.7 mmol) were added to 50 mL of methanol solution and stirred at room temperature for 30 min. Then, compound C4 (0.30 g, 1.4 mmol) was added and stirred at room temperature for 6 h. The mixture was then filtered, washed three times with methanol, and dried to obtain compound C5.

[0065] Compound C5 (0.18 g, 0.5 mmol) and malononitrile (0.03 g, 0.5 mmol) were added to 2-methoxyethanol (10 mL), and the mixture was heated under reflux for 18 h. The reaction solution was cooled to 5 °C, the precipitate was collected, washed with ethanol, and separated to obtain compound I-2.

[0066] Nuclear magnetic resonance and mass spectrometry, such as Figure 3 and Figure 4 As shown, Figure 3 This is a schematic diagram of the proton NMR spectrum of fluorescent probe I-2. Figure 4This is a schematic diagram of the high-resolution mass spectrum of fluorescent probe I-2. 1 H NMR (400MHz, DMSO-d6) δ8.75 (s, 1H), 8.62 (d, J = 8.4Hz, 1H), 7.82 (d, J = 7.9Hz ,1H),7.42-7.54(m,3H),3.47(s,4H),2.74-2.85(m,4H),1.91-1.97(m,4H). ESI-HRMS(m / z):[M+H] + Calculated for I-2(C 25 H 20 N5O,M + ):406.1668,found 406.1691.

[0067] Example 3

[0068] Fluorescence changes before and after the fluorescent probe reaction

[0069] 1. Fluorescence changes of fluorescent probe I-1 before and after reaction with different concentrations of nerve agents

[0070] Fluorescent probe I-1 was dissolved in dimethyl sulfoxide, and different concentrations of diethyl chlorophosphate (DCP) solution (0-100 μM) were added. The final concentration of fluorescent probe I-1 was 20 μM. After reacting for 1 minute, the solution was excited at 365 nm, and the fluorescence intensity was recorded at 582 nm. The results are as follows. Figure 5 As shown, Figure 5 This diagram illustrates the fluorescence changes of fluorescent probe I-1 before and after reacting with different concentrations of nerve agent. Fluorescent probe I-1 exhibits a strong fluorescence signal at 582 nm, but the fluorescence intensity at 582 nm gradually decreases with increasing nerve agent concentration. This indicates that the fluorescence of fluorescent probe I-1 is significantly quenched after reacting with the nerve agent diethyl chlorophosphate (DCP).

[0071] 2. Fluorescence changes of fluorescent probe I-1 before and after reaction with different concentrations of cyanide

[0072] Fluorescent probe I-1 was dissolved in dimethyl sulfoxide and then added to KCN solutions of different concentrations (0-200 μM). The final concentration of fluorescent probe I-1 was 20 μM. After reacting for 1 minute, the solution was excited at 365 nm, and the fluorescence intensity at 582 nm and 405 nm was recorded. The results are as follows: Figure 6 As shown, Figure 6This diagram illustrates the fluorescence changes of fluorescent probe I-1 before and after reacting with different concentrations of cyanide. Without cyanide, fluorescent probe I-1 exhibits a strong fluorescence signal at 582 nm and no fluorescence at 405 nm. After the addition of cyanide, the fluorescence intensity of fluorescent probe I-1 at 582 nm gradually decreases, and a new fluorescence emission peak appears at 405 nm, gradually increasing with increasing cyanide concentration. This indicates that the reaction of fluorescent probe I-1 with cyanide induces a blue shift in fluorescence.

[0073] 3. Fluorescence changes of fluorescent probe I-2 before and after reaction with different concentrations of nerve agents.

[0074] Fluorescent probe I-2 was dissolved in dimethyl sulfoxide, and different concentrations of DCP solution (0-100 μM) were added respectively, with a final concentration of 20 μM for the fluorescent probe. After reacting for 1 minute, the solution was excited at 365 nm, and the fluorescence intensity was recorded at 596 nm. The results are as follows. Figure 7 As shown, Figure 7 This diagram illustrates the fluorescence changes of fluorescent probe I-2 before and after reacting with different concentrations of nerve agent. Fluorescent probe I-2 exhibits a strong fluorescence signal at 596 nm, but the fluorescence intensity at 596 nm gradually decreases with increasing nerve agent concentration. This indicates that the fluorescence of fluorescent probe I-2 is significantly quenched after reacting with the nerve agent diethyl chlorophosphate (DCP).

[0075] 4. Fluorescence changes of fluorescent probe I-2 before and after reaction with different concentrations of cyanide

[0076] Fluorescent probe I-2 was dissolved in dimethyl sulfoxide and then added to KCN solutions of different concentrations (0-200 μM). The final concentration of the fluorescent probe was 20 μM. After reacting for 1 minute, the solution was excited at 365 nm, and the fluorescence intensity at 596 nm and 403 nm was recorded. The results are as follows: Figure 8 As shown, Figure 8 This diagram illustrates the fluorescence changes of fluorescent probe I-2 before and after reacting with different concentrations of cyanide. Fluorescent probe I-2 exhibits a strong fluorescence signal at 596 nm. As the cyanide concentration increases, the fluorescence intensity of fluorescent probe I-2 at 596 nm gradually decreases, while the fluorescence intensity at 403 nm gradually increases. This indicates that the reaction of fluorescent probe I-2 with cyanide induces a blue shift in fluorescence.

[0077] Example 4

[0078] Sensitivity and selectivity experiments

[0079] 1. Sensitivity

[0080] Based on the formula for calculating the detection limit of a probe, LOD = 3σ / k (where σ is the standard deviation of the target substance in the sample and k is the slope after fitting), fluorescent probe I-1 was dissolved in dimethyl sulfoxide, and different concentrations of DCP solution (final concentration 100 μM) were added. The final concentration of fluorescent probe I-1 was 20 μM. After reacting for 1 minute, the probe was excited at 365 nm, and the fluorescence intensity of the solution was recorded at 450 nm, confirming that the k value of the probe was 1.28. By detecting the background fluorescence of the probe 10 times, its σ was found to be 2.3, and the detection limit of fluorescent probe I-1 for DCP was 5.5 μM. Similarly, the detection limit of fluorescent probe I-1 for cyanide was found to be 9.6 nM.

[0081] The detection limit of fluorescent probe I-2 for DCP was 5.9 μM. Using the same method, the detection limit of fluorescent probe I-2 for cyanide was 10.6 μM.

[0082] 2. Reaction rate

[0083] Fluorescent probes I-1 and I-2 were dissolved in dimethyl sulfoxide, and nerve agent DCP (final concentration 100 μM) was added. The final concentrations of fluorescent probes I-1 and I-2 were 20 μM, respectively. Excitation was performed at 365 nm, and the changes in fluorescence intensity over 90 seconds were recorded at 582 nm or 596 nm. The results are as follows: Figure 9 As shown, Figure 9 This is a schematic diagram showing the fluorescence changes of fluorescent probes I-1 and I-2 at different time points after reacting with nerve agents. Both fluorescent probes I-1 and I-2 react with nerve agents within 10 seconds.

[0084] Fluorescent probes I-1 and I-2 were dissolved in dimethyl sulfoxide, and cyanide KCN (final concentration 200 μM) was added, resulting in final concentrations of 20 μM for both probes I-1 and I-2. Excitation was performed at 365 nm, and the changes in fluorescence intensity over 3 minutes were recorded at 582 nm / 596 nm and 405 nm / 403 nm. The results are as follows: Figure 10 As shown, Figure 10 This is a schematic diagram showing the fluorescence changes of fluorescent probes I-1 and I-2 reacting with cyanide at different times. Fluorescent probe I-1 reacts quickly, completing the reaction with cyanide within 1 minute, while fluorescent probe I-2 reacts with cyanide within 4 minutes.

[0085] 3. Selective Experiments

[0086] 3.1 Selectivity of fluorescent probe I-1 for nerve agents.

[0087] Fluorescent probe I-1 (20 μM) dissolved in dimethyl sulfoxide was added to different solutions containing or without nerve agents, numbered from 1 to 9: blank control, mustard gas, 2-chloroethyl sulfide, acetic acid, triphosgene, Lewisite, formaldehyde, diethylamine, and DCP. The fluorescence intensity of each solution was recorded. The results are as follows: Figure 11 As shown. Figure 11 This is a schematic diagram illustrating the selectivity of fluorescent probe I-1 for nerve agents. As shown in the figure, compared to the blank control, the fluorescence intensity of fluorescent probe I-1 is significantly reduced in the presence of only nerve agents. When substances 2-8 are present alone, the fluorescence intensity of fluorescent probe I-1 is not significantly different from the blank control group. However, once DCP is added, the fluorescence intensity significantly decreases, indicating that the fluorescent probe I-1 in this invention has high specificity and reacts only with nerve agents.

[0088] Like I-1, fluorescent probe I-2 exhibits good selectivity for nerve agents.

[0089] 3.2 Selectivity for cyanide

[0090] For cyanide fluorescent probes, the main interference affecting their selectivity is anions. Fluorescent probe I-1 (20 μM) dissolved in dimethyl sulfoxide is numbered from 1 to 9 as blank control, SCN, etc. - F - Cl - HPO4 2- HSO3 - Cys, GSH, and KCN were used as fluorescence spectra, which were recorded by a recorder. The results are as follows: Figure 12 As shown, Figure 12 This is a schematic diagram illustrating the selectivity of fluorescent probe I-1 for cyanide. As can be seen from the figure, KCN induces a significant fluorescence response in fluorescent probe I-1 at 582 nm, indicating that fluorescent probe I-1 also exhibits good selectivity for KCN.

[0091] Like I-1, fluorescent probe I-2 exhibits good selectivity for cyanides.

[0092] Example 5

[0093] The dual-target fluorescent probe I-1 of the present invention, as a reagent for the fluorescence detection of nerve agents, includes the following steps:

[0094] A dimethyl sulfoxide solution of the dual-target fluorescent probe I-1 was added to the sample solution to achieve a final concentration of 20 μM. After incubation for 1 min, the solution was excited at 365 nm, and the fluorescence intensity was recorded at 582 nm. Figure 13 As shown, Figure 13This is a schematic diagram of the detection of nerve agents and cyanide by fluorescent probes I-1 and I-2. A dual-target fluorescent probe solution is used as a blank control. If the sample contains nerve agents, the fluorescence intensity at 582 nm is weaker compared to the blank control.

[0095] The sample to be tested was selected from an organic solvent (acetonitrile) containing the nerve agent DCP.

[0096] The dual-target fluorescent probe I-2 of the present invention, as a reagent for the fluorescence detection of nerve agents, includes the following steps:

[0097] A dimethyl sulfoxide solution of the dual-target fluorescent probe I-2 was added to the sample solution to achieve a final concentration of 20 μM. After incubation for 1 min, the solution was excited at 365 nm, and the fluorescence intensity was recorded at 596 nm. Figure 13 As shown, using a dual-target fluorescent probe solution as a blank control, if the sample to be tested contains nerve agents, the fluorescence intensity at 596 nm is reduced compared to the blank control.

[0098] The sample to be tested was selected from an organic solvent (acetonitrile) containing the nerve agent DCP.

[0099] Example 6

[0100] The dual-target fluorescent probe I-1 of the present invention, used as a reagent for cyanide fluorescence detection, includes the following steps:

[0101] The dual-target fluorescent probe I-1 solution was added to the sample solution to make the final concentration of the dual-target fluorescent probe 20 μM. After incubation for 1 min, the solution was excited at 365 nm, and the fluorescence intensity at 582 nm and 405 nm, or the fluorescence intensity at 596 nm and 403 nm, was recorded.

[0102] like Figure 13 As shown, using a dual-target fluorescent probe solution as a blank control, if the sample contains cyanide, the fluorescence intensity at 582 nm is weaker than that of the blank control, while the fluorescence intensity at 405 nm is stronger.

[0103] The sample to be tested was selected from a solvent (methanol) containing KCN.

[0104] The dual-target fluorescent probe I-2 of the present invention, used as a reagent for cyanide fluorescence detection, includes the following steps:

[0105] The dual-target fluorescent probe I-2 solution was added to the sample solution to make the final concentration of the dual-target fluorescent probe 20 μM. After incubation for 1 min, the solution was excited at 365 nm, and the fluorescence intensity at 596 nm and 403 nm was recorded.

[0106] like Figure 13 As shown, using a dual-target fluorescent probe solution as a blank control, if the sample contains cyanide, the fluorescence intensity at 596 nm is weaker than that of the blank control, while the fluorescence intensity at 403 nm is stronger.

[0107] The sample to be tested was selected from a solvent (methanol) containing KCN.

[0108] Example 7

[0109] The cells used in the experiment were immortalized human epidermal cells (HaCaT cells). Cells were cultured in a standard cell culture incubator at 37°C and 5% CO2. Cells to be treated were seeded into wells 24 hours in advance. Before co-incubation with the probe, the culture medium was removed, and the probe was dissolved in fresh complete culture medium to a concentration of 20 μM before being added to the wells for co-incubation with the cells. Cells were divided into three groups: a blank group, a nerve agent group, and a cyanide group. All cells were incubated with I-1 for 10 min, followed by the addition of a nerve agent mimic, DCP or KCN. Before imaging experiments, the cells were carefully washed again with PBS buffer to remove the culture medium. Results are as follows: Figure 14 As shown, Figure 14 This is a schematic diagram of fluorescence imaging of live cells incubated with fluorescent probe I-1 reacting with different concentrations of nerve agents or cyanide. After incubation with cells, significant fluorescence is observed in the red channel, while no fluorescence is observed in the blue channel. Upon addition of the nerve agent mimic DCP, the fluorescence in the red channel gradually weakens, while the blue channel remains fluorescence-free. Upon addition of cyanide, fluorescence appears in the blue channel, while the fluorescence in the red channel gradually weakens.

[0110] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-described technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A dual-target fluorescent probe for detecting nerve agents and cyanide, characterized in that, The structure of the dual-target fluorescent probe for detecting nerve agents and cyanide is selected from the following structures: 。 2. The application of a dual-target fluorescent probe in the preparation of fluorescent detection reagents for nerve agents and / or fluorescent detection reagents for cyanide, characterized in that, The structure of the dual-target fluorescent probe is selected from one of the following structures: 。 3. The application according to claim 2, characterized in that, The application of the dual-target fluorescent probe as a reagent for the fluorescence detection of nerve agents includes the following steps: The dual-target fluorescent probe solution was added to the sample solution to achieve a final concentration of 20 μM. After incubation for 1 min, the sample was excited at 365 nm, and the fluorescence intensity of the solution was recorded at 582 nm or 596 nm. The dual-target fluorescent probe solution was used as a blank control. If the sample contained nerve agents, the fluorescence intensity at 582 nm or 596 nm was lower than that of the blank control. If the sample did not contain nerve agents, the fluorescence intensity at 582 nm or 596 nm remained unchanged compared with the blank control.

4. The application according to claim 2, characterized in that, The application of the dual-target fluorescent probe as a cyanide fluorescence detection reagent includes the following steps: The dual-target fluorescent probe solution was added to the sample solution to make the final concentration of the dual-target fluorescent probe 20 μM. After incubation for 1 min, the solution was excited at 365 nm, and the fluorescence intensity at 582 nm and 405 nm, or the fluorescence intensity at 596 nm and 403 nm, was recorded. Using the dual-target fluorescent probe solution as a blank control, if the sample contains cyanide, the fluorescence intensity at 582 nm is weaker than that of the blank control, while the fluorescence intensity at 405 nm is stronger; if the sample does not contain cyanide, the fluorescence intensity at 582 nm remains unchanged compared to the blank control, and the fluorescence intensity at 405 nm also remains unchanged compared to the blank control. Alternatively, a dual-target fluorescent probe solution can be used as a blank control. If the sample contains cyanide, the fluorescence intensity at 596 nm will be weaker than that of the blank control, while the fluorescence intensity at 403 nm will be stronger. If the sample does not contain cyanide, the fluorescence intensity at 596 nm will remain unchanged compared to the blank control, and the fluorescence intensity at 403 nm will also remain unchanged compared to the blank control.

5. The application according to claim 3, characterized in that, The samples to be tested are selected from organic solvents containing nerve agents, soil containing nerve agents, or living cells containing nerve agents.

6. The application according to claim 4, characterized in that, The samples to be tested were selected from solvents containing cyanide, soil containing cyanide, or living cells containing cyanide.

7. The application of a dual-target fluorescent probe in the preparation of fluorescent detection elements for nerve agents or cyanide, characterized in that, The structure of the dual-target fluorescent probe is selected from one of the following structures: 。 8. The application according to claim 7, characterized in that, The nerve agent fluorescence detection element or cyanide fluorescence detection element is selected from fluorescent test strips or kits.

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