An organic fluorescent probe for detecting tricyclazole and a preparation method and application thereof

Abstract

This invention discloses an organic fluorescent probe for detecting tricyclazole, its preparation method, and its application, belonging to the field of organic synthesis and analytical chemistry. The organic fluorescent probe uses cyanostilbene as the chromophore and is modified with (1R,2R)-1,2-cyclohexanediamine and a crown ether chain to obtain a fluorescent macrocyclic structure, i.e., the organic fluorescent probe, chemically named (1R,2R)-cyclo-tris(ethoxy)-bridged-di(4-phenyl(Z-1-cyanovinyl)phenyl)-4-oxoacetylcyclohexanediamine. The organic fluorescent probe of this invention exhibits good sensing ability for tricyclazole, strong anti-interference ability, and can achieve real-time, visual, and selective detection of tricyclazole. When tricyclazole is present in the environment, the organic fluorescent probe undergoes significant fluorescence quenching. The concentration of tricyclazole in the sample can be determined based on the good linear relationship between fluorescence intensity and the equivalent concentration of tricyclazole. Furthermore, the construction method of this organic fluorescent probe is efficient, and the detection steps are simple, making it an important reference in the field of agricultural detection.

Description

Technical Field

[0001] This invention belongs to the field of organic synthesis and analytical chemistry technology, specifically relating to an organic fluorescent probe for detecting tricyclazole, its preparation method, and its application. Background Technology

[0002] Organic fluorescent probes are substances in which an organic compound with a specific structure, under certain conditions, interacts with a analyte, resulting in changes in fluorescence signal such as increases or decreases in fluorescence intensity and shifts in emission peak position. Organic fluorescent probes enable qualitative or quantitative analysis of analytes by detecting the generated fluorescence. Due to their advantages such as tunable structure, high sensitivity, high selectivity, and visualization, they show great application potential in environmental analysis, biomarking, cell and tissue imaging, and clinical diagnosis and treatment. The design and synthesis of organic fluorescent probes with excellent fluorescence response to specific guest molecules or ions has attracted considerable attention from researchers and has broad application prospects.

[0003] Tricyclazole (5-methyl-1,2,4-triazole[3,4-b]benzothiazole), a widely used pesticide, possesses strong bactericidal activity and is an effective agent for controlling rice blast. However, studies have shown that tricyclazole has mutagenic and carcinogenic properties. Furthermore, due to steric hindrance of the benzene ring, hydrophobicity, and the stable structure of its three rings, tricyclazole exhibits weak biodegradability and high stability in water-soil systems. With the discharge of wastewater from pesticide industrial production and the widespread application of tricyclazole in agriculture, it has become a common residual pollutant in soil, surface water, and groundwater, posing a significant potential threat to human health and the ecological environment. Direct discharge of wastewater containing tricyclazole without treatment will cause serious environmental pollution. Therefore, developing a routine monitoring method that is efficient, simple to operate, highly sensitive, and capable of real-time detection of tricyclazole residues is of great practical significance and application value.

[0004] Current research on tricyclic azole detection is relatively limited, and no ideal and effective organic fluorescent probe for tricyclic azole detection has been reported in existing literature. Therefore, designing and synthesizing an organic fluorescent probe sensitive to tricyclic azole, especially one with good sensing capabilities and a unique recognition effect on tricyclic azole, is of particular importance.

[0005] The invention patent with publication number CN111208234A provides a rapid detection method for triazole fungicides in soil, achieving rapid quantitative detection of triazole fungicides in soil and overcoming problems such as background matrix interference, complex pretreatment and purification steps, and easy false positives. However, this invention is only applicable to triazoles in soil media and cannot detect triazole fungicides in other media. Furthermore, this method can simultaneously identify tricyclazole, triamcinolone, tebuconazole, hexaconazole, and propiconazole in soil, so the identification efficiency for tricyclazole is not high. (The last sentence appears to be a separate, unrelated statement.) Invention 97377A discloses a method for simultaneously determining the residues of 30 pesticides in agricultural products. The method involves purifying the pretreated agricultural product sample to prepare an instrumental solution, and then using a triple quadrupole liquid chromatography-mass spectrometry (LC-MS) instrument to simultaneously determine the residues of 30 pesticides in the agricultural product. This method is particularly suitable for the detection and analysis of large batches of agricultural products. However, before use, it is necessary to prepare an adsorption and purification reagent to reduce matrix effects, adsorb pigments and excess water, and to set the chromatographic conditions, mass spectrometry conditions and other measurement parameters of the triple quadrupole LC-MS instrument in advance to ensure that no target substances are lost. Summary of the Invention

[0006] To address the problems existing in the prior art, the present invention aims to provide an organic fluorescent probe for detecting tricyclazole, its preparation method, and its application. The organic fluorescent probe has good sensing ability for tricyclazole, strong anti-interference ability, and can realize real-time visual selective detection of tricyclazole. When tricyclazole is present in the environment, the organic fluorescent probe undergoes significant fluorescence quenching. The concentration of tricyclazole in the sample can be determined based on the good linear relationship between fluorescence intensity and the equivalent concentration of tricyclazole.

[0007] The technical solution of the present invention is as follows:

[0008] One objective of this invention is to provide an organic fluorescent probe for detecting tricyclazole, wherein the organic fluorescent probe has a fluorescent macrocyclic structure and its chemical structural formula is as follows:

[0009] The molecular formula is C 46 H 46 N4O8.

[0010] Furthermore, the organic fluorescent probe uses cyanostilbene as the chromophore and is modified by introducing (1R,2R)-1,2-cyclohexanediamine and crown ether chains.

[0011] Furthermore, the organic fluorescent probe was characterized by the following infrared spectrum: (KBr), v / cm -1 :3275(-CONH),2930(CH),2210(-CN),1605(C=N);

[0012] The 1H NMR spectrum characterization is as follows: (400MHz, CDCl3): δppm 7.67 (d, J = 8.0Hz, 4H, ArH), 7.40 (d, J = 8.0Hz, 4H, ArH), 6.91 (d, J = 8.0Hz, 4H, ArH), 6.82-6.86 (m, 8H, NH, ArH and CNC = CH), 4.40 (d, J = 13.0Hz, 2H, OCH2CO), 3.76-4.01 (m, 14H, OCH2 and CH), 3.61 (d, J = 12.0Hz, 2H, OCH2CO), 2.19-2.22 (m, 2H, CH2), 1.83 (bs, 2H, CH2), 1.32-1.45 (m, 4H, CH2);

[0013] The carbon NMR spectrum characterization is as follows: (100MHz, CDCl3) δ 168.86, 159.56, 158.51, 138.18, 131.77, 130.93, 130.80, 130.02, 126.84, 115.94, 115.29, 115.11, 70.98, 69.65, 67.65, 66.58, 53.58, 29.71, 24.62;

[0014] Mass spectrometry (m / s): Calculated value C 46 H 46 N4O8: 782.3316, Measured value: 782.9984 [M] + ],805.0043[M+Na + ],821.3077[M+K + ].

[0015] The second objective of this invention is to provide a method for preparing a tricyclic azole organic fluorescent probe, comprising the following steps:

[0016] (1) Weigh out triethylene glycol di-toluenesulfonate, K2CO3, KI and p-hydroxyphenylacetonitrile respectively and put them into a three-necked flask containing dry acetonitrile. Stir well at room temperature and start heating. During the process, use TLC to detect the reaction. When the raw materials react completely, stop heating. Add dilute hydrochloric acid while hot to remove K2CO3. After multiple extractions and rotary evaporation to remove the solvent, purify by column chromatography. Then, rotary evaporate and recrystallize again to obtain di-p-acetonitrile phenyl triethylene glycol.

[0017] (2) Weigh out di-p-acetonitrile-2-acetonitrile-vinyl)-phenyltriglyceride and p-hydroxybenzaldehyde respectively and put them into a three-necked flask. Weigh out NaOH and put it into a beaker containing anhydrous ethanol to dissolve and stir evenly. Then slowly pour it into the three-necked flask, heat and stir, and use TLC technology to detect the reaction during the process. After the reaction is complete, stop heating, add hydrochloric acid to neutralize the base until the solution is neutral, then add water and refrigerate to crystallize the product. Filter and dry to obtain di-4-(1-p-hydroxyphenyl-2-acetonitrile-vinyl)-phenyltriglyceride;

[0018] (3) Weigh (1R,2R)-1,2-cyclohexanediamine and chloroacetyl chloride (molar ratio 1:2) and add them to dichloromethane. Heat and stir under reflux. During the reaction, use TLC to monitor the reaction. After the reaction is complete, evaporate the solvent, add methanol, refrigerate to crystallize the product, filter and dry to obtain (1R,2R)-1,2-di-chloroacetamidocyclohexane.

[0019] (4) Under nitrogen protection, add di-4-(1-p-hydroxyphenyl-2-acetonitrile-vinyl)-phenyltriglyceride, anhydrous potassium carbonate, potassium iodide and (1R,2R)-1,2-dichloroacetamidocyclohexane to the reaction vessel, reflux in the dry acetonitrile system, and monitor the reaction by thin-layer chromatography.

[0020] (5) After the raw material disappears, add HCl solution dropwise to the reaction mixture, stir thoroughly, and then use chloroform to extract and separate the organic phase. Evaporate most of the solvent under reduced pressure, and separate the product by silica gel column chromatography to obtain (1R,2R)-cyclo-tris(ethoxy)-bridged-di(4-phenyl(Z-1-cyanovinyl)phenyl)-4-oxoacetylcyclohexanediamine, which is an organic fluorescent probe.

[0021] Furthermore, the synthesis methods of di-p-acetonitrile phenyl triglycan and di-4-(1-p-hydroxyphenyl-2-acetonitrile-vinyl)-phenyl triglycan were based on published literature (Ma Haifeng, Synthesis and Properties of Novel Cyanobrystyrene Fluorescent Probes, Master's Thesis, Fujian Normal University, 2022).

[0022] Furthermore, the synthesis method of (1R,2R)-1,2-dichloroacetamidocyclohexane was based on the literature, synthesized from (1R,2R)-1,2-cyclohexanediamine and chloroacetyl chloride (Zhang, X.; Jiang S.; Lin G.; Guo, H.; Yang, F. Novel fluorescent columnar liquid crystal based on

[0023] tetraphenylethylene-rufigallol-tetraphenylethylene triads, Journal of Molecular Structure, 2022, 1252, 132210).

[0024] Furthermore, in step (4), the molar ratio of di-4-(1-p-hydroxyphenyl-2-acetonitrile-vinyl)-phenyltriglyceride, anhydrous potassium carbonate, potassium iodide and (1R,2R)-1,2-dichloroacetamamidocyclohexane is 1:1~5:1~5:0.5~1.5.

[0025] Furthermore, the reflux time in step (4) is 12 to 48 hours.

[0026] Furthermore, the concentration of the HCl solution in step (5) is 0.8–1.2 M.

[0027] Furthermore, in step (5), the eluent for silica gel column chromatography is dichloromethane.

[0028] A third objective of this invention is to provide an application of an organic fluorescent probe for detecting tricyclazole. The organic fluorescent probe can be used for the sensitive detection of the pesticide tricyclazole in the environment. The concentration of tricyclazole in a sample can be determined based on a standard working curve showing the change in fluorescence intensity with the concentration of the pesticide tricyclazole.

[0029] Furthermore, the standard working curve establishment steps are as follows: prepare a THF / H2O solution containing the organic fluorescent probe, wherein the volume ratio of THF:H2O is 1:9; prepare a series of tricyclazole solutions with gradient concentrations according to different multiples of the THF / H2O solution; mix the THF / H2O solution with the tricyclazole series solutions respectively; measure the fluorescence intensity after mixing; and establish a standard working curve with fluorescence intensity on the ordinate and tricyclazole concentration on the abscissa.

[0030] Furthermore, the determination process involves mixing the THF / H2O solution containing the organic fluorescent probe with the sample to be tested, comparing the measured sample fluorescence intensity value with the standard working curve of the tricyclazole concentration gradient change, and reading the concentration of tricyclazole in the sample from the curve.

[0031] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0032] 1. This invention presents, for the first time, an organic fluorescent probe capable of quantitatively detecting tricyclazole, which has the following advantages: First, this organic fluorescent probe has high sensitivity; changes in tricyclazole content can cause significant quenching of the probe fluorescence, resulting in good analytical sensitivity. Second, it has strong anti-interference ability; other substances present in the sample have little impact on the fluorescence of this organic fluorescent probe, enabling it to produce a unique recognition effect on tricyclazole. Third, the detection results are visualized; this organic fluorescent probe is pale blue in tetrahydrofuran solution and exhibits weak fluorescence emission at 435 nm. In a tetrahydrofuran and water (1:9) solution, it exhibits strong cyan-green fluorescence emission at 475 nm. When it generates a strong intermolecular interaction with tricyclazole, this intermolecular interaction significantly weakens the fluorescence. Therefore, real-time, visualized, and selective detection of tricyclazole can be achieved based on the fluorescence signal.

[0033] 2. The organic fluorescent probe disclosed in this invention exhibits a good linear relationship between its fluorescence intensity and the equivalent concentration of tricyclazole. In the obtained standard working curve, Pearson'r = 1, R... 2 =1, Adj.R 2 =1. This organic fluorescent probe can simultaneously achieve qualitative and quantitative detection of tricyclazole, providing a better understanding of the distribution and dynamic changes of tricyclazole in samples. It can be widely used for routine monitoring of pesticide tricyclazole residues in environmental samples.

[0034] 3. Compared with the prior art, the organic fluorescent probe of the present invention uses cyanostilbene as the chromophore and introduces (1R,2R)-1,2-cyclohexanediamine and crown ether chain to modify it, thus preparing a novel fluorescent macrocyclic structure as a fluorescent probe for detecting tricyclazole. It has the advantages of simple synthesis method, wide availability of raw materials, and simple operation steps, providing a new material and method for convenient, rapid and highly sensitive determination of tricyclazole, and has good application prospects in the field of pesticide residue detection and analysis technology.

[0035] Figure Labels

[0036] Figure 1 This is a process flow diagram of the organic fluorescent probe preparation method described in this invention;

[0037] Figure 2 The infrared spectrum of the organic fluorescent probe described in this invention;

[0038] Figure 3 This is the proton NMR spectrum of the organic fluorescent probe described in this invention;

[0039] Figure 4 This is the carbon NMR spectrum of the organic fluorescent probe described in this invention;

[0040] Figure 5 This is the mass spectrum of the organic fluorescent probe described in this invention;

[0041] Figure 6 1×10 -5 A mol / L THF / H2O mixed solution containing the organic fluorescent probe of this invention and 1×10 -5 Fluorescence emission spectra of pesticide molecules at mol / L and metal ions that may exist in the water environment;

[0042] Figure 7 1×10 -5 Fluorescence spectra of a THF / H2O mixed solution containing the organic fluorescent probe of this invention at mol / L and tricyclazole at different concentrations;

[0043] Figure 8 This is a standard working curve showing the change in fluorescence intensity of the THF / H2O mixed solution containing an organic fluorescent probe and the change in tricyclazole concentration gradient in Example 3 of the present invention.

[0044] Figure 9 1×10 -5 A comparison of the fluorescence intensity ratios of the THF / H2O mixed solution containing the organic fluorescent probe of this invention in other guests (or other guests containing tricyclazole) to the fluorescence intensity of the THF / H2O mixed solution. Detailed Implementation

[0045] The present invention will be further described below with reference to the accompanying drawings and preferred embodiments. The embodiments given are only for illustrating the present invention and are not intended to limit the scope of the present invention.

[0046] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0047] In the quantitative experiments in the following examples, three replicate experiments were set up, and the average value of the results was taken.

[0048] Unless otherwise specified, the experimental methods described in the following examples are conventional methods.

[0049] Example 1

[0050] This embodiment provides an organic fluorescent probe for detecting tricyclazole, the preparation method of which includes the following steps:

[0051] (1) Weigh out triethylene glycol di-toluenesulfonate, K2CO3, KI and p-hydroxyphenylacetonitrile respectively and put them into a three-necked flask containing dry acetonitrile. Stir well at room temperature and start heating. During the process, use TLC to detect the reaction. When the raw materials react completely, stop heating. Add dilute hydrochloric acid while hot to remove K2CO3. After multiple extractions and rotary evaporation to remove the solvent, purify by column chromatography. Then, rotary evaporate and recrystallize again to obtain di-p-acetonitrile phenyl triethylene glycol.

[0052] (2) Weigh out di-p-acetonitrile-2-acetonitrile-vinyl)-phenyltriglyceride and p-hydroxybenzaldehyde respectively and put them into a three-necked flask. Weigh out NaOH and put it into a beaker containing anhydrous ethanol to dissolve and stir evenly. Then slowly pour it into the three-necked flask, heat and stir, and use TLC technology to detect the reaction during the process. After the reaction is complete, stop heating, add hydrochloric acid to neutralize the base until the solution is neutral, then add water and refrigerate to crystallize the product. Filter and dry to obtain di-4-(1-p-hydroxyphenyl-2-acetonitrile-vinyl)-phenyltriglyceride;

[0053] (3) Weigh (1R,2R)-1,2-cyclohexanediamine and chloroacetyl chloride (molar ratio 1:2) and add them to dichloromethane. Heat and stir under reflux. During the reaction, use TLC to monitor the reaction. After the reaction is complete, evaporate the solvent, add methanol, and refrigerate to crystallize the product. Filter and dry to obtain (1R,2R)-1,2-dichloroacetamidocyclohexane;

[0054] (4) Under nitrogen protection, 1.0 mmol of di-4-(1-p-hydroxyphenyl-2-acetonitrile-vinyl)-phenyltriglyceride, 1.0 mmol of anhydrous potassium carbonate, 1.0 mmol of potassium iodide and 1.5 mmol of (1R,2R)-1,2-dichloroacetamamidocyclohexane were added to a 250 mL three-necked flask in a molar ratio of 2:2:2:1. The mixture was refluxed in a dry acetonitrile system for 48 h, and the reaction was monitored by thin-layer chromatography.

[0055] (5) After the raw material disappears, add 1.2M HCl solution dropwise to the reaction mixture, stir thoroughly, and then extract the organic phase with chloroform. Evaporate most of the solvent under reduced pressure, and separate the product by silica gel column chromatography with dichloromethane as the eluent to obtain (1R,2R)-cyclo-tris(ethoxy)-bridged-di(4-phenyl(Z-1-cyanovinyl)phenyl)-4-oxoacetylcyclohexanediamine, i.e., the organic fluorescent probe, with a yield of about 54%.

[0056] Example 2

[0057] This embodiment provides a method for preparing an organic fluorescent probe for detecting tricyclazole, comprising the following steps:

[0058] (1) Weigh out triethylene glycol di-toluenesulfonate, K2CO3, KI and p-hydroxyphenylacetonitrile respectively and put them into a three-necked flask containing dry acetonitrile. Stir well at room temperature and start heating. During the process, use TLC to detect the reaction. When the raw materials react completely, stop heating. Add dilute hydrochloric acid while hot to remove K2CO3. After multiple extractions and rotary evaporation to remove the solvent, purify by column chromatography. Then, rotary evaporate and recrystallize again to obtain di-p-acetonitrile phenyl triethylene glycol.

[0059] (2) Weigh out di-p-acetonitrile-2-acetonitrile-vinyl)-phenyltriglyceride and p-hydroxybenzaldehyde respectively and put them into a three-necked flask. Weigh out NaOH and put it into a beaker containing anhydrous ethanol to dissolve and stir evenly. Then slowly pour it into the three-necked flask, heat and stir, and use TLC technology to detect the reaction during the process. After the reaction is complete, stop heating, add hydrochloric acid to neutralize the base until the solution is neutral, then add water and refrigerate to crystallize the product. Filter and dry to obtain di-4-(1-p-hydroxyphenyl-2-acetonitrile-vinyl)-phenyltriglyceride;

[0060] (3) Weigh (1R,2R)-1,2-cyclohexanediamine and chloroacetyl chloride (molar ratio 1:2) and add them to dichloromethane. Heat and stir under reflux. During the reaction, use TLC to monitor the reaction. After the reaction is complete, evaporate the solvent, add methanol, and refrigerate to crystallize the product. Filter and dry to obtain (1R,2R)-1,2-dichloroacetamidocyclohexane;

[0061] (4) Under nitrogen protection, 1.0 mmol of di-4-(1-p-hydroxyphenyl-2-acetonitrile-vinyl)-phenyltriglyceride, 5.0 mmol of anhydrous potassium carbonate, 5.0 mmol of potassium iodide and 0.5 mmol of (1R,2R)-1,2-dichloroacetamamidocyclohexane were added to a 250 mL three-necked flask in a molar ratio of 1:2:3:1. The mixture was refluxed in a dry acetonitrile system for 12 h, and the reaction was monitored by thin-layer chromatography.

[0062] (5) After the raw material disappears, add 0.8M HCl solution dropwise to the reaction mixture, stir thoroughly, and then extract the organic phase with chloroform. Evaporate most of the solvent under reduced pressure, and separate the product by silica gel column chromatography with dichloromethane as the eluent to obtain (1R,2R)-cyclo-tris(ethoxy)-bridged-di(4-phenyl(Z-1-cyanovinyl)phenyl)-4-oxoacetylcyclohexanediamine, i.e., the organic fluorescent probe, with a yield of about 46%.

[0063] Example 3

[0064] This embodiment provides an application of an organic fluorescent probe for detecting tricyclazole, comprising the following steps:

[0065] (1) Weigh out triethylene glycol di-toluenesulfonate, K2CO3, KI and p-hydroxyphenylacetonitrile respectively and put them into a three-necked flask containing dry acetonitrile. Stir well at room temperature and start heating. During the process, use TLC to detect the reaction. When the raw materials react completely, stop heating. Add dilute hydrochloric acid while hot to remove K2CO3. After multiple extractions and rotary evaporation to remove the solvent, purify by column chromatography. Then, rotary evaporate and recrystallize again to obtain di-p-acetonitrile phenyl triethylene glycol.

[0066] (2) Weigh out di-p-acetonitrile-2-acetonitrile-vinyl)-phenyltriglyceride and p-hydroxybenzaldehyde respectively and put them into a three-necked flask. Weigh out NaOH and put it into a beaker containing anhydrous ethanol to dissolve and stir evenly. Then slowly pour it into the three-necked flask, heat and stir, and use TLC technology to detect the reaction during the process. After the reaction is complete, stop heating, add hydrochloric acid to neutralize the base until the solution is neutral, then add water and refrigerate to crystallize the product. Filter and dry to obtain di-4-(1-p-hydroxyphenyl-2-acetonitrile-vinyl)-phenyltriglyceride;

[0067] (3) Weigh (1R,2R)-1,2-cyclohexanediamine and chloroacetyl chloride (molar ratio 1:2) and add them to dichloromethane. Heat and stir under reflux. During the reaction, use TLC to monitor the reaction. After the reaction is complete, evaporate the solvent, add methanol, and refrigerate to crystallize the product. Filter and dry to obtain (1R,2R)-1,2-dichloroacetamidocyclohexane;

[0068] (4) Under nitrogen protection, 1.0 mmol of di-4-(1-p-hydroxyphenyl-2-acetonitrile-vinyl)-phenyltriglyceride, 3.0 mmol of anhydrous potassium carbonate, 3.0 mmol of potassium iodide and 1.0 mmol of (1R,2R)-1,2-dichloroacetamamidocyclohexane were added to a 250 mL three-necked flask in a molar ratio of 2:10:10:3. The mixture was refluxed in a dry acetonitrile system for 24 h, and the reaction was monitored by thin-layer chromatography.

[0069] (5) After the raw material disappears, add 1M HCl solution dropwise to the reaction mixture, stir thoroughly, and then extract the organic phase with chloroform. Evaporate most of the solvent under reduced pressure, and separate the product by silica gel column chromatography with dichloromethane as the eluent to obtain (1R,2R)-cyclo-tris(ethoxy)-bridged-di(4-phenyl(Z-1-cyanovinyl)phenyl)-4-oxoacetylcyclohexanediamine, i.e., the organic fluorescent probe, with a yield of about 86%.

[0070] (6) Prepare a THF / H2O solution containing the above-mentioned organic fluorescent probe, wherein the volume ratio of THF:H2O is 1:9. Prepare a series of tricyclic azole solutions with gradient concentrations of 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 2.0, 3.0, 4.0, and 5.0 times the concentration of the organic fluorescent probe. Mix the THF / H2O solution with each of these tricyclic azole solutions, measure the fluorescence intensity after mixing, and establish a standard working curve with fluorescence intensity on the ordinate and tricyclic azole concentration on the abscissa, as shown below. Figure 8 As shown;

[0071] (7) Mix the organic fluorescent probe with the tricyclazole solution to be tested, compare the measured fluorescence intensity value with the standard working curve of tricyclazole concentration gradient change, and read the tricyclazole concentration in the tricyclazole solution to be tested from the curve.

[0072] like Figure 6 As shown, when different guest tests were performed, the THF / H2O mixed solution containing the organic fluorescent probe of the present invention exhibited a strong interaction with the presence of tricyclazole, and the fluorescence was significantly quenched, indicating that this organic fluorescent probe has good selective detection capability for tricyclazole.

[0073] exist Figure 7 As the concentration of tricyclazole continued to increase, the fluorescence intensity of the THF / H2O mixed solution containing the organic fluorescent probe of the present invention showed a significant decreasing trend. Therefore, it can be inferred that there is a good linear relationship between the fluorescence intensity of the mixed solution containing the organic fluorescent probe and the equivalent concentration of tricyclazole, which can be applied to quantitative analysis activities related to tricyclazole.

[0074] Figure 9 In the middle, I is 1×10 -5 mol / L organic fluorescent probe and 1×10 -5 Fluorescence intensity of a mixture of other guests (or other guests + tricyclazole) at mol / L, I o The fluorescence intensity is represented by a 1×10⁻⁵ mol / L organic fluorescent probe. From... Figure 9 It can be seen that the fluorescence ratios of the organic fluorescent probes are all close to 1 after the addition of other guests, indicating that the other guests have little effect on the fluorescence of the organic fluorescent probes. However, after adding tricyclazole to the other guests, the fluorescence of the organic fluorescent probes weakens, with the I / I0 ratio around 0.18, indicating that the other guests have little interference with the sensitive detection of tricyclazole by the fluorescent probes of this invention.

[0075] The above description is merely an embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structural or procedural transformations made based on the content of the present invention specification, or direct or indirect applications in other related technical fields, are similarly included within the patent protection scope of the present invention.

Claims

1. An organic fluorescent probe for detecting tricyclazole, characterized in that, The organic fluorescent probe is a fluorescent macrocyclic structure, and its chemical structural formula is: The molecular formula is C 46 H 46 N4O8.

2. A method for preparing the organic fluorescent probe for detecting tricyclazol according to claim 1, characterized by, The method comprises the following steps: (1) under the protection of nitrogen, adding di-4-(1-p-hydroxyphenyl-2-acetonitrile-ethenyl)-phenyl triglycol, anhydrous potassium carbonate, potassium iodide and (1R,2R)-1,2-dichloroacetamidocyclohexane in a reaction container, refluxing in a dry acetonitrile system, and monitoring the reaction by thin layer chromatography; (2) after the raw materials disappear, adding an HCl solution into the reaction mixture, stirring sufficiently, extracting and separating the organic phase by using chloroform, evaporating most of the solvent under reduced pressure, and separating the product by using a silica gel column chromatography to obtain (1R,2R)-cyclo- tris(ethoxy)-bridged-di(4-phenyl(Z-1-cyanovinyl)phenyl)-4-oxyacetylcyclohexanediamine, i.e. the organic fluorescent probe.

3. The method for preparing an organic fluorescent probe for detecting tricyclazole as described in claim 2, characterized in that, In the step (1), the molar ratio of di-4-(1-p-hydroxyphenyl-2-acetonitrile-ethenyl)-phenyl triglycol, anhydrous potassium carbonate, potassium iodide and (1R,2R)-1,2-dichloroacetamidocyclohexane is 1:1-5:1-5:0.5-1.

5.

4. The method for preparing an organic fluorescent probe for detecting tricyclazole as described in claim 2, characterized in that, In the step (1), the refluxing time is 12-48 h.

5. The method for preparing an organic fluorescent probe for detecting tricyclazole as described in claim 2, characterized in that, In the step (2), the concentration of the HCl solution is 0.8-1.2 M.

6. The method for preparing an organic fluorescent probe for detecting tricyclazole as described in claim 2, characterized in that, In the step (2), the eluent for the silica gel column chromatography separation is dichloromethane.

7. An application of the organic fluorescent probe in the sensitive detection of tricyclazole according to claim 1.

8. The use of an organic fluorescent probe according to claim 7, characterized in that, The THF / H2O solution of the organic fluorescent probe is added into a sample solution to be detected, and the tricyclazole in the sample solution can be sensitively detected by comparing the standard working curve of the change of the fluorescence intensity with the tricyclazole concentration.

9. The use of an organic fluorescent probe according to claim 8, characterized in that, In the THF / H2O solution, the volume ratio of THF to H2O is 1:9.

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