Nopinic alkyl acridine double-detection fluorescent probe for detecting hydrazine and viscosity as well as preparation method and application of nopinic alkyl acridine double-detection fluorescent probe

Abstract

The invention discloses a nopinic alkyl acridine fluorescent probe for detecting hydrazine and viscosity as well as a preparation method and application of the nopinic alkyl acridine fluorescent probe. The probe is 2-(2-(5-(4-(3, 3-dimethyl-1, 2, 3, 4-tetrahydro-2, 4-methylene acridine-9-yl) phenyl) furan-2-yl) vinyl)-1, 1, 3-trimethyl-1H-benzo [e] indoleonium iodized salt, which is called TA-PFV-BI for short. The probe can detect the viscosity of a solution, and red fluorescence emitted by the probe is gradually enhanced along with the improvement of the viscosity of the solution under the irradiation of ultraviolet light with the wavelength of 365 nm. The probe can also have a specific reaction with hydrazine, so that the fluorescence color of a probe solution is changed from colorless to bright blue, the detection range is 0-90 mu M, the response time is within 5 seconds, the detection limit is 4 * 10 <-8 > M, and the applicable pH range is 5-10. The probe TA-PFV-BI, as a bifunctional fluorescent probe capable of simultaneously detecting hydrazine and viscosity, has the advantages of good selectivity, high sensitivity, fast response, wide application range and the like, and shows a good application prospect.

Description

Technical Field

[0001] This invention belongs to the field of fluorescent probe technology, and relates to a nopinel acridine dual-detection fluorescent probe for detecting hydrazine and viscosity, its preparation method and application. Background Technology

[0002] Hydrazine (N₂H₄) is an important chemical raw material widely used in military, chemical, pharmaceutical, and pesticide industries. As a strong reducing agent, it can be used for metallizing plastics and glass surfaces; in the military field, it is used as fuel for jet engines and rockets; it is an important raw material for the synthesis of drugs such as aminourea, furacilin, and isoniazid; and it can also be used as a foaming agent in the preparation of foam materials. However, hydrazine is a toxic substance that can easily cause poisoning through skin contact and ingestion. It strongly corrodes the skin and damages the eyes and liver. Hydrazine is also a mutagen and carcinogen. It easily leads to severe pollution of the atmosphere, water, and soil, posing a serious threat to human health. Furthermore, changes in intracellular viscosity are closely related to various pathological processes; for example, neurodegenerative diseases, diabetes, and cancer all cause increased intracellular viscosity. Real-time monitoring of cellular microenvironment viscosity has become an important tool for disease diagnosis. Therefore, developing a multifunctional fluorescent probe capable of simultaneously detecting hydrazine and monitoring changes in cellular viscosity is of great significance for environmental safety and biomedical research.

[0003] Fluorescent probes, due to their high sensitivity, excellent spatiotemporal resolution, and non-invasive detection characteristics, have become important tools for bioimaging and environmental analysis. Despite significant progress in the research and application of fluorescent probes, developing multifunctional fluorescent probes remains a challenge. To address the shortcomings of existing fluorescent probes for detecting hydrazine and viscosity, such as long response times, low sensitivity, narrow detection range, high cytotoxicity, and the limitation to single detection, this invention aims to develop a bifunctional fluorescent probe for detecting hydrazine and viscosity that features fast response time, high sensitivity, wide detection range, and applicability to cell imaging. Summary of the Invention

[0004] To address the shortcomings of existing technologies, the technical problem to be solved by this invention is to develop a nopinel acridine fluorescent probe that can simultaneously detect hydrazine and viscosity. This probe has the characteristics of fast response, high sensitivity, and wide pH adaptability. It can be used for qualitative and quantitative detection of hydrazine in water, plants, and aquatic animals, and can be applied to fluorescence imaging of hydrazine and viscosity in cells.

[0005] To solve the above-mentioned technical problems, the technical solution adopted by this invention is as follows: A norpinyl acridine-based fluorescent probe for detecting hydrazine and viscosity is developed, with the following structural formula:

[0006]

[0007] The fluorescent probe has the molecular formula C0.43 H 39 IN2O, chemically named 2-(2-(5-(4-(3,3-dimethyl-1,2,3,4-tetrahydro-2,4-bridged methylene acridine-9-yl)phenyl)furan-2-yl)vinyl)-1,1,3-trimethyl-1H-benzo[e]indoline iodide, abbreviated as TA-PFV-BI.

[0008] The method for preparing the norpinyl acridine fluorescent probe for detecting hydrazine and viscosity involves reacting 5-(4-(3,3-dimethyl-1,2,3,4-tetrahydro-2,4-bridged methylene acridine-9-yl)phenyl)furan-2-carboxaldehyde with 1,1,2,3-tetramethyl-1H-benzo[e]indolium iodide to obtain the compound TA-PFV-BI. Specifically, the method includes the following steps:

[0009] (1) Add 0.25 mmol, 98.4 mg of 5-(4-(3,3-dimethyl-1,2,3,4-tetrahydro-2,4-bridged methylene acridine-9-yl)phenyl)furan-2-carboxaldehyde, 0.25-1.00 mmol, 87.8-351.2 mg of 1,1,2,3-tetramethyl-1H-benzo[e]indoline iodide salt and 10-50 mL of anhydrous ethanol to a reaction flask. Slowly add 0.25-1.00 mmol, 13.5-54.02 mg of sodium ethoxide at 60-70 °C. Stir the reaction for 10-30 min, then heat to reflux and continue the reaction for 10-20 h.

[0010] (2) After the reaction is completed, the ethanol is recovered by distillation. The distillation residue is dissolved in 30-50 mL of toluene, washed with water and saturated saline until neutral, dried with anhydrous sodium sulfate and then the toluene is evaporated to obtain the crude product TA-PFV-BI.

[0011] (3) The crude product of TA-PFV-BI was recrystallized with a mixed solvent of dichloromethane and petroleum ether to obtain red powder TA-PFV-BI.

[0012] The compound TA-PFV-BI specifically reacts with hydrazine. Under 365 nm UV light irradiation, the probe solution changes from colorless to bright blue fluorescence upon the addition of hydrazine. The linear detection range for hydrazine is 0-90 μM, with a response time within 5 s and a detection limit of 0.04 μM. The applicable pH range is 5-10. TA-PFV-BI also specifically identifies solution viscosity. Under 365 nm UV light irradiation, the red fluorescence of the probe significantly increases with increasing solution viscosity. The linear detection range for viscosity is 1.28 cP-1411.98 cP, with a response time within 1 s and a detection limit of 1.00 cP.

[0013] Beneficial Effects: Compared with existing technologies, this invention uses 5-(4-(3,3-dimethyl-1,2,3,4-tetrahydro-2,4-bridged methylene acridine-9-yl)phenyl)furan-2-carboxaldehyde as a raw material, reacting it with 1,1,2,3-tetramethyl-1H-benzo[e]indoline iodide to prepare the compound TA-PFV-BI. This compound can specifically identify hydrazine and viscosity, and can sensitively detect the content of hydrazine in a solution as well as the viscosity of the solution. As a bifunctional fluorescent probe for detecting hydrazine and viscosity, it has many advantages such as good selectivity, high sensitivity, and wide application range, and has good application prospects. Attached Figure Description

[0014] Figure 1 The fluorescence spectra of TA-PFV-BI reacting with different concentrations of hydrazine are shown.

[0015] Figure 2 The fluorescence spectra of TA-PFV-BI reacting with solvents of different viscosities are shown.

[0016] Figure 3 This is a bar chart showing the fluorescence intensity at 450 nm of TA-PFV-BI interacting with different analytes, such as metal ions, anions, and common amino acids.

[0017] Figure 4 These are fluorescence spectra of TA-PFV-BI reacting with different solvents. Detailed Implementation

[0018] The present invention will be further described below with reference to specific embodiments.

[0019] Example 1

[0020] The preparation of TA-PFV-BI is shown in the following reaction formula:

[0021]

[0022] The specific steps are as follows:

[0023] Preparation of TA-PFV-BI:

[0024] 98.4 mg of 5-(4-(3,3-dimethyl-1,2,3,4-tetrahydro-2,4-bridged methylene acridine-9-yl)phenyl)furan-2-carboxaldehyde, 87.8 mg of 1,1,2,3-tetramethyl-1H-benzo[e]indoline iodide, and 15 mL of anhydrous ethanol were added to a reaction flask. 13.5 mg of sodium ethoxide was added at 60 °C, and the mixture was stirred for 10 min. The temperature was then slowly increased, and the reaction was continued under reflux for 10 h. After the reaction was complete, the ethanol was recovered by distillation. The distillation residue was dissolved in 30–50 mL of toluene, washed with water and saturated brine until neutral, dried with anhydrous sodium sulfate, and the toluene was evaporated. The crude TA-PFV-BI product was recrystallized from the dichloromethane-petroleum ether mixture to obtain the compound TA-PFV-BI, a red powder. 1 H NMR (600MHz, DMSO-d6) δ: 8.49-8.43 (m, 2H), 8.36-8.26 (m, 3H), 8.22 (d, J = 7.9Hz, 1H), 8.11 (d, J = 8.9Hz, 1H), 7.97 (d, J =8.3Hz, 1H), 7.83-7.79 (m, 1H), 7.74 (d, J = 3.8Hz, 1H), 7.71 (d, J = 6.9Hz, 1H), 7.68 (t, J = 8.3Hz, 1H), 7.61 (d, J = 10.3Hz, 1H), 7.59 (d, J=3.7Hz, 1H), 7.54 (d, J=8.6Hz, 1H), 7.45 (d, J=16.0Hz, 2H), 7.39-7.34 (m, 1H), 4.26 (s, 3H), 3.14 (d, J=5 .6Hz, 1H), 2.83 (d, J=17.6Hz, 2H), 2.03 (s, 6H), 1.88 (s, 1H), 1.44 (s, 3H), 1.43-1.37 (m, 2H), 1.23 (s, 1H), 0.71 (s, 3H).

[0025] Example 2

[0026] The compound TA-PFV-BI was prepared into 1×10 -5 Simultaneously, hydrazine was dissolved in PBS buffer solution to prepare concentrations of 0 and 1 × 10⁻⁶ mol / L (pH = 7.4, 1‰ Tween 80). -7 2×10 -7 3×10 -7 4×10 -7 5×10 -7 6×10 -7 7×10 -7 8×10 -7 9×10 -7A mol / L solution. The fluorescence emission spectra of TA-PFV-BI in the presence of different concentrations of hydrazine were measured using a fluorescence spectrophotometer via fluorescence spectrophotometric titration. For example... Figure 1 As shown in the figure. The results indicate that as the concentration of hydrazine in the solution gradually increases from 0 mol / L to 9 × 10⁻⁶, the effect is more pronounced. -7 At a concentration of mol / L, the fluorescence emission intensity of this compound gradually increases at 450 nm. This indicates that the compound can be used as an on-type fluorescent probe for the sensitive detection of hydrazine.

[0027] Example 3

[0028] TA-PFV-BI was made into 1×10 -3 A 0.1 mol / L PBS buffer solution (pH = 7.4) was prepared, and glycerol solutions of different concentrations (0%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 99.9%) were also dissolved in the PBS buffer. 100 μL of TA-PFV-BI PBS buffer solution was added to 10 mL of the glycerol solution, and the fluorescence emission spectra of TA-PFV-BI in the glycerol solutions of different concentrations were measured using a fluorescence spectrophotometer. Figure 2 As shown, the fluorescence emission intensity of this compound at 650 nm gradually increases as the glycerol concentration in the solution gradually increases from 0 to 100%. This indicates that the compound can be used as a fluorescent probe for sensitive detection of solution viscosity.

[0029] Example 4

[0030] TA-PFV-BI was formulated into 1×10 -5 A 100 μM solution of TA-PFV-BI was prepared by dissolving various analytes, including metal ions, anions, and common amino acids, in PBS buffer (pH = 7.4, 1‰ Tween 80). Fluorescence emission spectra of the TA-PFV-BI in the presence of these analytes were measured using a fluorescence spectrophotometer via fluorescence titration. Figure 3 As shown, the compound exhibited a distinct fluorescence emission peak at 450 nm after reacting with hydrazine, while the addition of Fe... 3+ Cu 2+ Mg 2+ Ca 2+ Zn 2+ Ni 2+ Metal ions, F - I - ,Br - HS - NO 2-Compared with anions and various analytes such as Tyr, Gln, and Cys amino acids, the fluorescence spectrum of this compound did not show significant changes. This indicates that this compound can be used as an open-type fluorescent probe for the selective detection of hydrazine.

[0031] Example 5

[0032] TA-PFV-BI was made into 1×10 -3 Prepare a 100 μL PBS buffer solution (pH = 7.4), along with glycerol and other organic solvents. Add 100 μL of TA-PFV-BI in PBS buffer solution to 10 mL of the solvent. Measure the fluorescence emission spectra of TA-PFV-BI in different solvents using a fluorescence spectrophotometer, as shown below. Figure 4 As shown, the compound exhibits a distinct fluorescence emission peak at 650 nm in glycerol solution, while no significant change occurs in other solvents such as water, dimethyl sulfoxide, and tetrahydrofuran. This indicates that the compound can be used as an open-type fluorescent probe for detecting solution viscosity.

Claims

1. A norpinane-based acridinium fluorescent probe, characterized in that, The fluorescent probe is 2-(2-(5-(4-(3,3-dimethyl-1,2,3,4-tetrahydro-2,4-bridged methylene acridine-9-yl) phenyl) furan-2-yl) vinyl)-1,1,3-trimethyl-1H-benzo[e] indolium iodide, which is abbreviated as TA-PFV-BI, and its structural formula is:

2. The method for preparing the norpinane-based acridinium fluorescent probe according to claim 1, characterized in that, 5-(4-(3,3-dimethyl-1,2,3,4-tetrahydro-2,4-bridged methylene acridine-9-yl) phenyl) furan-2-carboxaldehyde is used as a raw material to react with 1,1,2,3-tetramethyl-1H-benzo[e] indolium iodide to obtain the compound TA-PFV-BI.

3. The method for preparing the nopinel-based acridine fluorescent probe according to claim 2, characterized in that, The specific preparation method comprises the following steps: (1) 0.25 mmol, 98.4 mg of 5-(4-(3,3-dimethyl-1,2,3,4-tetrahydro-2,4-bridged methylene acridine-9-yl) phenyl) furan-2-carboxaldehyde, 0.25-1.00 mmol, 87.8-351.2 mg of 1,1,2,3-tetramethyl-1H-benzo[e] indolium iodide and 10-50 mL of anhydrous ethanol are added into a reaction bottle, 0.25-1.00 mmol, 13.5-54.02 mg of sodium ethoxide is slowly added at 60-70 DEG C, the reaction is stirred for 10-30 min, and then heating is performed to reach a reflux state, and the reaction is continuously performed for 10-20 h; (2) after the reaction is completed, the reaction liquid is subjected to distillation to recover ethanol, the distillation residue is dissolved in 30-50 mL of toluene, washed with water and saturated brine until neutral, dried with anhydrous sodium sulfate, and then toluene is evaporated to obtain the crude product of TA-PFV-BI; (3) the crude product of TA-PFV-BI is subjected to recrystallization with a dichloromethane-petroleum ether mixed solvent to obtain red powder TA-PFV-BI.

4. The application of the no pinane acridine fluorescent probe in solution viscosity and hydrazine detection according to claim 1.

5. Use according to claim 4, characterized in that: The probe TA-PFV-BI can specifically react with hydrazine, under the irradiation of a 365 nm ultraviolet lamp, the probe solution is changed from no light to bright blue fluorescence after the addition of hydrazine, the linear detection range of the probe to hydrazine is 0-90 μM, the response time is within 5 s, the detection limit is 0.04 μM, and the applicable pH range is 5-10.

6. Use according to claim 4, characterized in that: The probe TA-PFV-BI can also specifically identify solution viscosity, under the irradiation of a 365 nm ultraviolet lamp, the red fluorescence of the probe is significantly enhanced with the increase of the solution viscosity. The linear detection range of the probe to viscosity is 1.28 cP-1411.98 cP, the response time is within 1 s, and the detection limit is 1.00 cP.

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