Fluorescence probe rapidly responding to hydrogen sulfide and preparation method and application thereof

Abstract

The invention discloses a fluorescence probe rapidly responding to hydrogen sulfide and a preparation method and application thereof. The structural formula of the compound can be seen in the specification. The preparation method includes the step of making hydroxystyrene and 2,4-dinitrosalicylic bromobenzene react under the conditions of catalysts and heating to generate the compound named as TPE-NP. Probe molecules have large Stokes shift, self-absorption is effectively reduced, and imaging accuracy is improved. In addition, the fluorescence probe is quite sensitive to hydrogen sulfide, and fluorescence is enhanced by 40 times in the process of increasing the concentration of sodium bisulfide to 10 equivalent weights of the probe concentration from 0. The fluorescence probe has a high molar extinction coefficient (larger than 60000 M<-1>.CM<-1> except for water), has a good dyeing effect on living cells and nematode, and is short in dyeing time (15 min) and high in dyeing efficiency. After living cells and nematode are dyed, confocal imaging is directly conducted without conducting the washing process which must be conducted on other probes in the prior art, and the imaging operation process is greatly simplified.

Description

technical field

[0001] The invention belongs to the field of synthesis and technology, and in particular relates to a fluorescent probe that responds quickly to hydrogen sulfide and its preparation method and application. Background technique

[0002] Hydrogen sulfide gas is well known as a toxic molecule, but recent studies have found that it can be used as a signal molecule for life activities such as anti-oxidation, ischemia-reperfusion injury, antibacterial and anti-inflammatory, and apoptosis. Hydrogen sensors are extremely important. Although there are a variety of detection methods for hydrogen sulfide, most of them are based on the principles of electrochemistry, gas chromatography, etc., and cannot detect hydrogen sulfide quickly, in situ, and visually.

[0003] Fluorescence imaging technology has developed into the most important means to dynamically monitor biomolecular events in living cells and in vivo. The development of new fluorescent imaging sensors has gr...

Examples

Embodiment 1

[0042] In a 25ml two-necked flask, add hydroxytetraphenylethylene (0.364g, 1mmol), 2,4-dinitrobromobenzene (0.738g, 3mmol), activated potassium carbonate (0.414g, 3mmol), and vacuum— - Nitrogen was passed three times, water-removed DMF was added, the temperature was raised to 90° C., and the reaction was carried out for 4 hours. The solution turned yellow. After the reaction, the reaction liquid was poured into the ice-water mixture, the product was precipitated rapidly, and the mixed liquid was milky, extracted with a large amount of ethyl acetate, and the ethyl acetate phase was a yellow liquid. Add anhydrous magnesium sulfate to dry. Magnesium sulfate was filtered off, spin-dried, and passed through the column. The eluent is 5 to 1 petroleum ether to dichloromethane, R f The value is about 0.25. After being passed through the column and spin-dried, a light yellow solid (0.341 g, 0.49 mmol) was obtained with a yield of 49%.

Embodiment 2

[0044] In a 25ml two-necked flask, add hydroxytetraphenylethylene (1mmol), 2,4-dinitrobromobenzene (2mmol), activated potassium carbonate (0.414g, 3mmol), vacuumize---pass nitrogen three times, add DMF was removed, the temperature was raised to 85°C, and the reaction was carried out for 5 hours. The solution turned yellow. After the reaction, the reaction liquid was poured into the ice-water mixture, the product was precipitated rapidly, and the mixed liquid was milky, extracted with a large amount of ethyl acetate, and the ethyl acetate phase was a yellow liquid. Add anhydrous magnesium sulfate to dry. Magnesium sulfate was filtered off, spin-dried, and passed through the column. The eluent is 5 to 1 petroleum ether to dichloromethane, R f The value is about 0.25. After passing through the column, it was spin-dried to obtain a light yellow solid with a yield of 45%.

Embodiment 3

[0046] In a 25ml two-neck flask, add hydroxytetraphenylethylene (1mmol), 2,4-dinitrobromobenzene (4mmol), activated potassium carbonate (3mmol), vacuumize --- pass nitrogen three times, add dehydrated DMF , heated up to 95°C, and reacted for 4 hours. The solution turned yellow. After the reaction, the reaction liquid was poured into the ice-water mixture, the product was precipitated rapidly, and the mixed liquid was milky, extracted with a large amount of ethyl acetate, and the ethyl acetate phase was a yellow liquid. Add anhydrous magnesium sulfate to dry. Magnesium sulfate was filtered off, spin-dried, and passed through the column. The eluent is 5 to 1 petroleum ether to dichloromethane, R f The value is about 0.25. After passing through the column, it was spin-dried to obtain a light yellow solid with a yield of 42%.

[0047] Characterization of melting point, infrared, NMR and mass spectrometry:

[0048] Probe melting point (m.p.):224-225℃

[0049] 1 HNMR (400MHz...

technical field

[0001] The invention belongs to the field of synthesis and technology, and in particular relates to a fluorescent probe that responds quickly to hydrogen sulfide and its preparation method and application. Background technique

[0002] Hydrogen sulfide gas is well known as a toxic molecule, but recent studies have found that it can be used as a signal molecule for life activities such as anti-oxidation, ischemia-reperfusion injury, antibacterial and anti-inflammatory, and apoptosis. Hydrogen sensors are extremely important. Although there are a variety of detection methods for hydrogen sulfide, most of them are based on the principles of electrochemistry, gas chromatography, etc., and cannot detect hydrogen sulfide quickly, in situ, and visually.

[0003] Fluorescence imaging technology has developed into the most important means to dynamically monitor biomolecular events in living cells and in vivo. The development of new fluorescent imaging sensors has gr...

Examples

Embodiment 1

[0042] In a 25ml two-necked flask, add hydroxytetraphenylethylene (0.364g, 1mmol), 2,4-dinitrobromobenzene (0.738g, 3mmol), activated potassium carbonate (0.414g, 3mmol), and vacuum— - Nitrogen was passed three times, water-removed DMF was added, the temperature was raised to 90° C., and the reaction was carried out for 4 hours. The solution turned yellow. After the reaction, the reaction liquid was poured into the ice-water mixture, the product was precipitated rapidly, and the mixed liquid was milky, extracted with a large amount of ethyl acetate, and the ethyl acetate phase was a yellow liquid. Add anhydrous magnesium sulfate to dry. Magnesium sulfate was filtered off, spin-dried, and passed through the column. The eluent is 5 to 1 petroleum ether to dichloromethane, R f The value is about 0.25. After being passed through the column and spin-dried, a light yellow solid (0.341 g, 0.49 mmol) was obtained with a yield of 49%.

Embodiment 2

[0044] In a 25ml two-necked flask, add hydroxytetraphenylethylene (1mmol), 2,4-dinitrobromobenzene (2mmol), activated potassium carbonate (0.414g, 3mmol), vacuumize---pass nitrogen three times, add DMF was removed, the temperature was raised to 85°C, and the reaction was carried out for 5 hours. The solution turned yellow. After the reaction, the reaction liquid was poured into the ice-water mixture, the product was precipitated rapidly, and the mixed liquid was milky, extracted with a large amount of ethyl acetate, and the ethyl acetate phase was a yellow liquid. Add anhydrous magnesium sulfate to dry. Magnesium sulfate was filtered off, spin-dried, and passed through the column. The eluent is 5 to 1 petroleum ether to dichloromethane, R f The value is about 0.25. After passing through the column, it was spin-dried to obtain a light yellow solid with a yield of 45%.

Embodiment 3

[0046] In a 25ml two-neck flask, add hydroxytetraphenylethylene (1mmol), 2,4-dinitrobromobenzene (4mmol), activated potassium carbonate (3mmol), vacuumize --- pass nitrogen three times, add dehydrated DMF , heated up to 95°C, and reacted for 4 hours. The solution turned yellow. After the reaction, the reaction liquid was poured into the ice-water mixture, the product was precipitated rapidly, and the mixed liquid was milky, extracted with a large amount of ethyl acetate, and the ethyl acetate phase was a yellow liquid. Add anhydrous magnesium sulfate to dry. Magnesium sulfate was filtered off, spin-dried, and passed through the column. The eluent is 5 to 1 petroleum ether to dichloromethane, R f The value is about 0.25. After passing through the column, it was spin-dried to obtain a light yellow solid with a yield of 42%.

[0047] Characterization of melting point, infrared, NMR and mass spectrometry:

[0048] Probe melting point (m.p.):224-225℃

[0049] 1 HNMR (400MHz...

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