A flavonol aromatic ether light-up fluorescent probe for detecting hydrogen sulfide, and its preparation method and application

Abstract

The present invention discloses a flavonol aromatic ether light-up type fluorescent probe for detecting hydrogen sulfide and its preparation method and application. The present invention uses 7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)-3-hydroxychromone as raw material, and carries out etherification reaction with 2,4-dinitrofluorobenzene to prepare a flavonol aromatic ether light-up type fluorescent probe. The flavonol aromatic ether light-up type fluorescent probe can react with hydrogen sulfide quickly and exclusively at room temperature. Under 365nm ultraviolet light, the solution fluorescence color changes from colorless to orange-red, the minimum detection limit is 96nM, and the response time is 3min. Therefore, the flavonol aromatic ether light-up type fluorescent probe has good application prospects in detecting hydrogen sulfide.

Description

Technical Field

[0001] The invention belongs to the technical field of fine organic synthesis and relates to a flavonol aromatic ether light-up type fluorescent probe for detecting hydrogen sulfide, and a preparation method and application thereof. Background Art

[0002] Hydrogen sulfide (H2S) plays a crucial role in food safety and living systems. It is found in fruits, vegetables, and meats rich in organic sulfur. Appropriate levels of H2S can extend the shelf life of fruits and vegetables by increasing the activity of antioxidant enzymes. However, excessive H2S production is also a sign of food spoilage. Protein-rich foods, including meat, seafood, and eggs, produce excessive amounts of H2S during decay and spoilage. Furthermore, H2S has been reported to be one of the factors that affects the quality of red wine and deteriorates its taste. Therefore, high concentrations of H2S can degrade food quality and potentially harm human health. Previously, H2S was considered an irritating, colorless, and highly toxic gas. Further research has revealed that endogenous H2S functions similarly to nitric oxide and carbon monoxide, acting as a gaseous transmitter in the body for signaling. H2S has numerous physiological functions, such as anti-inflammatory and anti-tumor activities, regulating neurotransmission, and inhibiting insulin signaling. However, abnormal levels of H2S in cells can lead to various diseases, such as Alzheimer's disease, Down syndrome, diabetes, and cirrhosis. Therefore, it is of great significance to develop a method for hydrogen sulfide detection with high sensitivity and selectivity.

[0003] Numerous methods have been reported for detecting hydrogen sulfide, including high-performance liquid chromatography, gas chromatography, colorimetry, and electrochemical analysis. However, these methods are limited by complex sample pretreatment, expensive equipment, cumbersome procedures, and time-consuming detection. In contrast, fluorescent probes offer advantages such as high sensitivity, rapid response, high selectivity, real-time analysis, good biocompatibility, and non-invasiveness, allowing for real-time imaging of living cells. Organic small molecule fluorescent probes, among others, have recently become an important method for detecting hydrogen sulfide due to their small size, simple synthesis, fast reaction time, good selectivity, high sensitivity, and the ability to use fluorescence as an output signal for real-time detection. Summary of the Invention

[0004] In response to the deficiencies in the prior art, the technical problem to be solved by the present invention is to provide a flavonol aromatic ether light-up type fluorescent probe for detecting hydrogen sulfide, which can react rapidly and specifically with hydrogen sulfide at room temperature. Under 365nm ultraviolet light, the solution changes from colorless to orange-red fluorescence, allowing qualitative and quantitative detection of hydrogen sulfide. Another technical problem to be solved by the present invention is to provide a method for preparing a flavonol aromatic ether light-up type fluorescent probe for detecting hydrogen sulfide. Another technical problem to be solved by the present invention is to provide an application of a flavonol aromatic ether light-up type fluorescent probe for detecting hydrogen sulfide.

[0005] In order to solve the above technical problems, the technical solution adopted by the present invention is:

[0006] The flavonol aromatic ether light-emitting fluorescent probe is 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone, and its structural formula is:

[0007]

[0008] The preparation method of the flavonol aromatic ether light-up fluorescent probe is characterized in that the specific preparation method of 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone comprises:

[0009] (1) 5 mmol of 7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)-3-hydroxychromone, 5-15 mmol of 2,4-dinitrofluorobenzene, and 20-40 mL of DMF were added to a dry three-necked flask in sequence and reacted at room temperature for 1 h. 10-20 mmol of sodium carbonate was dissolved in 2-8 mL of deionized water and added to the reaction flask, and the reaction continued at room temperature for 24-30 h.

[0010] (2) Saturated brine was added to the reaction solution, and then extracted with ethyl acetate three times. The organic phase was washed with saturated brine until neutral, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product of 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone;

[0011] (3) The crude product of 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone was separated on a silica gel column (ethyl acetate / petroleum ether = 1 / 10, v / v) to obtain 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone as a white solid.

[0012] 3-(2,4-Dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone is used in the detection of hydrogen sulfide. It reacts rapidly and specifically with hydrogen sulfide at room temperature. Under 365nm ultraviolet light, the solution changes from colorless to orange-red fluorescent. The minimum detection limit is 96nM, and the response time is 3 minutes.

[0013] The flavonol aromatic ether light-up type fluorescent probe is used in detecting hydrogen sulfide.

[0014] Beneficial effects: Compared with the existing technology, the flavonol aromatic ether light-emitting fluorescent probe prepared using furfural, a natural renewable resource, as a raw material can react specifically with hydrogen sulfide. Under 365nm ultraviolet light, the fluorescent color of the solution changes from colorless to orange-red. It has good specificity, high sensitivity and fast response speed, and has good application prospects as a fluorescent probe for detecting hydrogen sulfide. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 is GSH, Cys, Hcy, Val, Gly, Arg, Thr, Leu, CH3COO - , SO3 2- , HSO3 - ,HCO3 - , CO3 2- , Cl - , Br - , NO3 - , I - , NO 2- , H2O2. etc., etc., on the fluorescence spectrum of 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone;

[0016] Figure 2 This is a graph showing the effect of different concentrations of hydrogen sulfide on the fluorescence spectrum of 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone;

[0017] Figure 3 This is a result graph of the response time spectrum of hydrogen sulfide to 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone. DETAILED DESCRIPTION

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

[0019] Example 1

[0020] Preparation of 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone, the reaction formula is as follows:

[0021]

[0022] 5mmol 7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)-3-hydroxychromone, 10mmol 2,4-dinitrofluorobenzene and 20-40mL DMF were added to a dry three-necked flask in sequence and reacted at room temperature for 1h; then 10mmol sodium carbonate was dissolved in deionized water and added to the reaction flask and reacted at room temperature for 24-30h; saturated brine was added to the reaction solution, and then extracted with ethyl acetate three times, the organic phase was washed with saturated brine until neutral, and then dried over anhydrous sodium sulfate, filtered and concentrated to obtain 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-( The crude product of 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone was separated by silica gel column (ethyl acetate / petroleum ether = 1 / 10, v / v) to obtain 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone as a white solid with a yield of 82.3% and a purity of 97.5%. 1 H NMR (600MHz, DMSO-d6) δ: 8.93 (d, J=2.8Hz, 1H), 8.34 (dd, J=9.3, 2.8Hz, 1H), 8.15 (d, J=1.6Hz, 1H), 8.08-8.04 (m, 2H), 7.87-7.80 (m, 3 H), 7.66 (d, J=9.3Hz, 1H), 7.43 (d, J=3.6Hz, 1H), 7.40-7.34 (m, 4H), 7.16-7.09 (m, 6H), 7.07-7.02 (m, 2H), 6.86 (dd, J=3.7, 1.7Hz, 1H). 13 C NMR (150MHz, DMSO-d6) δ: 170.59, 155.91, 153.93, 149.10, 148.82, 148.35, 147.11, 146.26, 142.90, 142.18, 138.74, 132 .33, 130.96, 130.24, 129.72, 128.79, 126.12, 125.40, 124.44, 124.06, 122.44, 122.26, 122.21, 118.33, 115.12, 114.01. 13C NMR (150MHz, Chloroform-d) δ170.54, 155.55, 154.28, 149.04, 147.34, 147.16, 146.84, 142.82, 141.84, 138.63, 132.14, 131.28, 12 9.50, 128.81, 128.11, 126.37, 125.13, 124.31, 123.79, 122.77, 122.31, 121.74, 118.38, 117.14, 114.98, 113.29.HRMS (m / z): [M+H] + calcd for C 37 H 23 N3O8+H + , 638.1563; found: 638.1511.

[0023] Example 2

[0024] 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone and GSH, Cys, Hcy, Val, Gly, Arg, Thr, Leu, CH3COO - , SO3 2- , HSO3 - ,HCO3 - , CO3 2- , Cl - , Br - , NO3 - , I - , NO 2- , H2O2 and other 19 different analytes were dissolved in DMSO / PBS (v / v=5 / 5) buffer solution to a concentration of 1.0×10 -6 M probe solution and the concentration was 1.0 × 10 -5 The fluorescence spectra of 19 different analyte solutions of M were measured by fluorescence spectrometer. The results are as follows Figure 1 shown. Figure 1 It shows that the addition of hydrogen sulfide changes the fluorescence intensity of the system from colorless to orange-red under 365nm ultraviolet light, while the addition of other interfering substances such as GSH, Cys, Hcy, Val, Gly, Arg, Thr, Leu, CH3COO - , SO3 2- , HSO3 - ,HCO3 - , CO3 2- , Cl - , Br - , NO3 -, I - , NO2 - , H2O2, etc., the fluorescence of the solution did not change, indicating that the compound can be used as a fluorescent probe to specifically identify hydrogen sulfide.

[0025] Example 3

[0026] 3-(2,4-Dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone was prepared to a concentration of 1.0×10 -5 M DMSO / PBS (v / v = 5 / 5) buffer solution, and hydrogen sulfide was similarly dissolved in DMSO / PBS (v / v = 5 / 5) buffer solution to prepare solutions with concentrations of 0, 10, 20, 30, 40, 50, 60, 70, 80, 90, and 100 μM. The effect of the fluorescence spectrum of 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone on the response to different concentrations of hydrogen sulfide was measured by titration. The results are shown in FIG. Figure 2 shown. Figure 2 It shows that as the concentration of hydrogen sulfide in the system continues to increase, the fluorescence intensity at 590nm gradually increases, which shows that the compound can sensitively detect hydrogen sulfide with a minimum detection limit of 96nM.

[0027] Example 4

[0028] 3-(2,4-Dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone was prepared to a concentration of 1.0×10 -5 M DMSO / PBS (v / v=5 / 5) buffer solution, and hydrogen sulfide was dissolved in DMSO / PBS (v / v=5 / 5) buffer solution to prepare a solution with a concentration of 100μM. The response time fluorescence spectrum of the solution was measured using a fluorescence spectrometer, and the results were as follows Figure 3 shown. Figure 3 The results showed that after adding hydrogen sulfide, the fluorescence intensity at the emission wavelength of 590nm gradually increased over time, reaching a maximum within 3 minutes and remaining stable. This shows that the compound can quickly detect hydrogen sulfide with a response time of 3 minutes.

Claims

1. A flavonol aromatic ether light-up type fluorescent probe for detecting hydrogen sulfide, characterized in that: The structural formula is: 。 2. The method for preparing the flavonol aromatic ether light-up type fluorescent probe according to claim 1, characterized in that: 7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)-3-hydroxychromone was used as the raw material and etherified with 2,4-dinitrofluorobenzene to prepare 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone.

3. The method for preparing the flavonol aromatic ether light-up type fluorescent probe according to claim 2, wherein: The following steps are involved: 1) 5 mmol of 7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)-3-hydroxychromone, 5-15 mmol of 2,4-dinitrofluorobenzene, and 20-40 mL of DMF were added sequentially to a dry three-necked flask and reacted at room temperature for 1 hour. 10-20 mmol of sodium carbonate was then dissolved in 2-8 mL of deionized water and added to the reaction flask, and the reaction continued at room temperature for 24-30 hours. 2) Saturated brine was added to the reaction solution, and then extracted with ethyl acetate three times. The organic phase was washed with saturated brine until neutral, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product of 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone; 3) The crude product of 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone was separated on a silica gel column with ethyl acetate / petroleum ether = 1 / 10, v / v, to give 3-(2,4-dinitrophenoxy)-7-(4-(diphenylamino)phenyl)-2-(furan-2-yl)chromone as a white solid.

4. Use of the flavonol aromatic ether light-up type fluorescent probe according to claim 1 as a fluorescent probe in detecting hydrogen sulfide.

5. The use according to claim 4, characterized in that: The flavonol aromatic ether light-emitting fluorescent probe can react with hydrogen sulfide at room temperature. Under 365nm ultraviolet light, the solution changes from colorless to orange-red fluorescence.

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