CN113307771B - Fluorescent dye with large Stokes displacement and preparation method thereof - Google Patents
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Abstract
The invention discloses a fluorescent dye with large Stokes displacement and a preparation method thereof, wherein isophorone and malononitrile are used as raw materials, and an intermediate TEM is synthesized through Knoevenagel condensation reaction; then, 4-bromo-1, 8-naphthalene dicarboxylic anhydride and N- (2-aminoethyl) morpholine are used as raw materials to synthesize an intermediate BMD; then, BMD and 4-piperazine-1-benzaldehyde are taken as raw materials to synthesize an intermediate NAA; finally, TEM and NAA are used as raw materials to synthesize the fluorescent dye. In the dye, naphthalimide is used as FRET energy donor, and dicyanoisophorone derivative is used as energy acceptor, so that Stokes displacement is increased. By testing the fluorescence emission spectra in various organic solvents, it was found that: stokes displacement is between 173 and 234nm, and has obvious fluorescence signal emission.
Description
Technical Field
The invention belongs to the technical field of fluorescent dye preparation, and particularly relates to a fluorescent dye with large Stokes displacement and a preparation method of the fluorescent dye.
Background
Traditional organic fluorescent dyes, such as BODIPY, fluorescein, rhodamine and the like, are commonly applied to the high and new technical fields of luminescent materials, fluorescent probes, fluorescent labels and the like. However, these organic fluorescent dyes have limited their wide application due to their drawbacks in terms of photophysical and optical properties during practical application.
Stokes shift (Stokes shift) is one of the important parameters for assessing the photophysical properties of fluorescent dyes. If the stokes shift of the organic dye itself is relatively small, fluorescence self-quenching is easily caused in the biological imaging process, thereby reducing the imaging resolution. Therefore, the reasonable design and synthesis of organic fluorescent dyes with large stokes shift have important research significance.
Disclosure of Invention
It is an object of the present invention to provide a fluorescent dye with a large Stokes shift, which is constructed based on fluorescence resonance energy transfer effect (FRET).
It is another object of the present invention to provide a method for preparing the above fluorescent dye with large Stokes shift.
The technical scheme adopted by the invention is that the fluorescent dye with large Stokes displacement has a structural formula shown in the following formula (I):
the preparation method of the fluorescent dye with large Stokes displacement is implemented according to the following steps:
and step 4, synthesizing the fluorescent dye by taking the intermediate TEM obtained in the step 1 and the intermediate NAA obtained in the step 3 as raw materials.
The present invention is also characterized in that,
in step 1, specifically:
dissolving isophorone, malononitrile and a catalyst in N, N-dimethylformamide, taking argon as a protective gas, stirring for 6 hours at 120 ℃, cooling to room temperature after the reaction is finished, injecting a reaction mixed solution into ice water, separating out brown solid, drying, and separating and purifying by using column chromatography to obtain an intermediate TEM; the catalyst is a viscous liquid formed by mixing acetic anhydride, glacial acetic acid and piperidine; the molar ratio of isophorone to malononitrile is 1:1.
In step 2, specifically:
dissolving 4-bromo-1, 8-naphthalene dicarboxylic anhydride and N- (2-aminoethyl) morpholine in 1, 4-dioxane, and reacting at 80 ℃ for 8 hours; cooling to room temperature after finishing, injecting the reaction solution into ice water to generate precipitate, filtering to obtain light yellow solid, drying, separating and purifying by batch column chromatography, wherein the eluent is dichloromethane to obtain white solid, namely intermediate BMD; the molar ratio of 4-bromo-1, 8-naphthalene dicarboxylic anhydride to N- (2-aminoethyl) morpholine was 1:2.
In step 3, specifically:
dissolving BMD, 4-piperazine-1-benzaldehyde and a catalyst in dimethyl sulfoxide, heating and stirring at 100 ℃, tracking by TLC until the reaction is finished, cooling to room temperature, adding water into a reaction liquid to generate yellow solid precipitate, carrying out suction filtration and air drying, and separating and purifying by using column chromatography to finally obtain a yellow-green solid product, namely an intermediate NAA; the molar ratio of BMD to 4-piperazine-1-benzaldehyde is 2.5:3, and cesium carbonate is used as a catalyst.
In step 4, specifically:
dissolving an intermediate TEM, an intermediate NAA and a catalyst in absolute ethyl alcohol, taking argon as a protective gas, heating and stirring for 4 hours at 80 ℃, cooling to room temperature after the reaction is finished, filtering to obtain a reddish brown solid product, and naturally airing to obtain the fluorescent dye with large Stokes displacement; the molar ratio of TEM to NAA is 2:3; the catalyst is piperidine.
The invention has the beneficial effects that fluorescent dye molecules with larger Stokes displacement are constructed by utilizing a fluorescence resonance energy transfer mechanism (FRET) and bridging by naphthalimide and dicyanoisophorone. The dye molecule of the invention has good strong acid resistance, is suitable for a wide pH range of neutrality and weak alkalinity, and is not easy to be interfered by other active ions. The fluorescent dye has the excellent performance and the application potential of imaging systems with different viscosities.
Drawings
FIG. 1 is a graph of fluorescence emission spectra of fluorescent dye TB (10. Mu. Mol/L) in various organic solutions;
FIG. 2 is DMSO H 2 pH stability test of fluorescent dye TB (10. Mu. Mol/L) in O (1:1, V:V) solution;
FIG. 3 is DMSO: H 2 Testing the interference performance of the fluorescent dye TB (10 mu mol/L) in O (1:1, V:V) solution against other active ions;
FIG. 4 is a graph showing the change in fluorescence spectrum of fluorescent dye TB (10. Mu. Mol/L) at various viscosities;
FIG. 5 is a schematic diagram of the design of a fluorescent dye prepared by the method of the present invention;
FIG. 6 is a diagram showing the reaction scheme of a fluorescent dye prepared in the example of the present invention.
Detailed Description
The invention will be described in detail below with reference to the drawings and the detailed description.
The invention discloses a fluorescent dye with large Stokes displacement, wherein fluorescent dye molecules have a structural formula shown in the following formula (I):
the invention discloses a preparation method of a fluorescent dye with large Stokes displacement, which is implemented according to the following steps:
the method comprises the following steps: dissolving isophorone, malononitrile and a catalyst in N, N-dimethylformamide, taking argon as a protective gas, stirring for 6 hours at 120 ℃, cooling to room temperature after the reaction is finished, injecting a reaction mixed solution into ice water, separating out brown solid, drying, and separating and purifying by using column chromatography to obtain an intermediate TEM;
the catalyst is a viscous liquid formed by mixing acetic anhydride, glacial acetic acid and piperidine; the molar ratio of isophorone to malononitrile is 1:1;
the method comprises the following steps: dissolving 4-bromo-1, 8-naphthalene dicarboxylic anhydride and N- (2-aminoethyl) morpholine in 1, 4-dioxane, and reacting at 80 ℃ for 8 hours; cooling to room temperature after finishing, injecting the reaction solution into ice water to generate precipitate, filtering to obtain light yellow solid, drying, separating and purifying by batch column chromatography, wherein the eluent is dichloromethane to obtain white solid, namely intermediate BMD;
the molar ratio of the 4-bromo-1, 8-naphthalene dicarboxylic anhydride to the N- (2-aminoethyl) morpholine is 1:2;
(Ⅳ)；
the method comprises the following steps: dissolving BMD, 4-piperazine-1-benzaldehyde and a catalyst in dimethyl sulfoxide, heating and stirring at 100 ℃, tracking by TLC until the reaction is finished, cooling to room temperature, adding a small amount of water into the reaction liquid to generate yellow solid precipitate, carrying out suction filtration and air drying, and separating and purifying by using column chromatography to finally obtain a yellow-green solid product, namely an intermediate NAA;
the molar ratio of BMD to 4-piperazine-1-benzaldehyde is 2.5:3, and the catalyst is cesium carbonate;
the method comprises the following steps: dissolving an intermediate TEM, an intermediate NAA and a catalyst in absolute ethyl alcohol, taking argon as a protective gas, heating and stirring for 4 hours at 80 ℃, cooling to room temperature after the reaction is finished, filtering to obtain a reddish brown solid product, and naturally airing to obtain a fluorescent dye TB with large Stokes displacement;
the molar ratio of TEM to NAA is 2:3; the catalyst is piperidine.
The design principle of the fluorescent dye prepared by the method is shown in figure 5;
fluorescence Resonance Energy Transfer (FRET) is an effective mechanism to increase Stokes shift. However, reasonable construction of fluorescent molecules with FRET mechanism is harsh. Mainly comprises three aspects, namely: 1) The two types of fluorophores involved must be at a suitable spatial distance; 2) The emission spectrum of a fluorescent molecule must overlap partially with the absorption spectrum of another fluorescent molecule; 3) A suitable refractive index. Therefore, through reasonable selection, the naphthalimide and the dicyanoisophorone derivative are bridged to obtain the fluorescent dye TB with excellent FRET effect and large Stokes displacement.
Examples
The preparation method of the fluorescent dye with large Stokes displacement is implemented according to the following steps, and the reaction formula is shown in figure 6;
the method comprises the following steps: to a 250mL three-necked flask under argon, 0.2g of acetic anhydride, 0.4mL of glacial acetic acid and 1.8mL of piperidine were added, followed by 16.5mL (110 mmol) of isophorone, 6.6g (110 mmol) of malononitrile and dissolved in 55mL of N, N-dimethylformamide, and stirred at 120℃for 6h. After the reaction was completed, the reaction mixture was cooled to room temperature. Pouring the reaction mixed solution into ice water, separating out brown solid, drying, separating and purifying by using column chromatography, wherein the eluent is petroleum ether, namely dichloromethane=1:1 (v: v), so as to obtain yellow solid;
the method comprises the following steps: 2.5g (9.0 mmol) of 4-bromo-1, 8-naphthalenedicarboxylic anhydride and N- (2-aminoethyl) morpholine were dissolved in 50ml of 1, 4-dioxane and refluxed overnight at 80℃and cooled to room temperature after completion. Thereafter, the reaction solution was poured into ice water to form a precipitate, and the precipitate was suction-filtered to obtain a pale yellow solid. Drying, separating and purifying by batch column chromatography, wherein the eluent is methylene dichloride, and finally obtaining white solid;
wherein the product is characterized as follows:
1 H NMR(400MHz,Chloroform-d)δ8.65(d,J＝7.2Hz,1H),8.57 (d,J＝8.5Hz,1H),8.40(d,J＝7.9Hz,1H),8.04(d,J＝7.8Hz,1H), 7.85(t,J＝7.9Hz,1H),4.39-4.27(m,2H),3.67(s,4H),2.70(t,J＝6.7 Hz,2H),2.59(s,4H).
13 C NMR(400MHz,Chloroform-d)δ163.70,133.38,132.12, 131.31,131.21,130.71,130.40,129.09,128.19,123.13,122.26,67.15, 56.20,53.93,37.45.
the method comprises the following steps: into a 25mL round bottom flask under argon, 1.0g (2.5 mmol) of BMD, 0.6g (3 mmol) of 4-piperazine benzaldehyde and 0.82g (3 mmol) of cesium carbonate were dissolved in 5mL of DMSO and heated to 100℃with stirring. TLC monitored the progress of the reaction and cooled to room temperature after completion. A small amount of water was added to the reaction solution, and a yellow solid was precipitated. Suction filtration, air drying, separation and purification by column chromatography, eluting agent: petroleum ether: dichloromethane = 1:1, 0.26g of a yellowish green solid product was finally obtained in 21% yield.
Wherein the product is characterized as follows:
1 H NMR(400MHz,Chloroform-d)δ9.84(s,1H),8.64-8.43(m, 3H),7.87-7.69(m,3H),7.28(d,J＝8.1Hz,1H),7.04(d,J＝8.9Hz, 2H),4.36(t,J＝6.9Hz,2H),3.79-3.64(m,8H),3.47-3.37(m,4H), 2.70(d,J＝39.2Hz,6H).
13 C NMR(400MHz,Chloroform-d)δ190.58,164.43,163.96, 155.36,154.97,132.55,131.96,131.36,130.01,127.92,126.23,123.36, 117.42,115.28,114.11,66.95,56.19,53.80,52.77,47.62,37.02.
the method comprises the following steps: under argon, 0.10g (0.2 mmol) NAA and 0.056g (2.0 mmol) TEM were dissolved in 5mL absolute ethanol, then 100. Mu.L piperidine was added and the reaction solution was heated to reflux and reacted overnight. After the reaction is finished, cooling to room temperature, filtering to obtain a reddish brown solid product, naturally airing, and finally obtaining 0.034g of solid product with 54% yield.
Wherein the product is characterized as follows:
1 H NMR(400MHz,Chloroform-d)δ8.59(d,J＝7.3Hz,1H),8.53 (d,J＝8.0Hz,1H),8.46(d,J＝8.4Hz,1H),7.72(td,J＝8.4,7.9,2.6Hz, 1H),7.48(d,J＝8.8Hz,1H),7.28(s,1H),7.20(d,J＝8.4Hz,1H),7.06 -6.97(m,2H),6.95-6.88(m,1H),6.87-6.81(m,1H),6.79(s,1H), 4.34(d,J＝6.7Hz,2H),3.78-3.65(m,4H),3.64-3.52(m,4H),3.43(s, 4H),2.74(s,2H),2.66-2.61(m,2H),2.58(s,1H),2.52(s,1H),2.46(s, 1H),2.23(s,1H),1.25(d,J＝3.7Hz,2H),1.07(s,3H),0.97(s,3H), 0.87(s,2H).
the fluorescence emission performance of the fluorescent dye TB prepared in this example was tested as follows:
fluorescence performance tests in different organic solvents.
The excitation wavelength was 450nm and fluorescence emission of fluorescent dye TB (10. Mu. Mol/L) in various organic solvents was tested, including: methanol (MeOH), acetonitrile (MeCN), ethanol (EtOH), ethyl Acetate (EA), dimethyl sulfoxide (DMSO), N-Dimethylformamide (DMF), dichloromethane (DCM), chloroform (CHCl) 3 ) Acetone (Acetone), toluene (tolene) and Tetrahydrofuran (THF). As a result, as shown in FIG. 1, it was found that there was a clear fluorescent signal emission in various organic solvents. And has larger Stokes displacement, which is between 173 and 234 nm.
pH stability test of fluorescent dye TB.
In DMSO: H 2 The stability of the fluorochromes in O (5:5, V:V) solutions over different pH ranges (2-11) was tested and shown in FIG. 2. The discovery is as follows: the fluorescent dye itself emits a distinct fluorescent signal in a strongly acidic solvent and has a good, readily observable fluorescent signal in a pH range (2-9) that includes neutral and weakly basic. Therefore, the fluorescent dye has a wide pH application range.
And (3) testing the anti-active ion interference performance of the fluorescent dye TB.
In DMSO: H 2 In O (5:5, V:V) solution, fluorescence spectra of different interfering ions added to fluorescent dye TB (10. Mu. Mol/L) were tested, including: 200. mu mol/L, 1) H 2 O 2 ；2)TBHP；3)NO 2 - ；4)Met；5)Hcy；6)GSH；7)HSO 3 2- ； 8)SO 3 2- ；9)HOCl；10)HS - . As shown in fig. 3, the fluorescence spectrum did not change significantly at the 450nm excitation wavelength, except for the three thiol analytes. The results show that the fluorescent dye TB can still maintain the excellent fluorescent property in the presence of various other interfering ions. In addition, when the viscosity of the solvent system is increased, such as glycerol (Gly), the fluorescence intensity value is obviously increased, and the application potential of the imaging system with different viscosities is shown.
Fluorescent dye TB was used for fluorescence effect testing in different viscosity systems.
In glycerol (Gly) and aqueous solution, the viscosity of the system is regulated by changing the proportion of the glycerol (Gly) and the aqueous solution. As shown in fig. 4, the fluorescence intensity value also showed a significant trend of increasing with increasing Gly ratio to 100%, i.e., increasing viscosity of the system. Further demonstrates the potential of fluorescent dyes for imaging different viscosity systems. The reason for this phenomenon may be due to the presence of double bonds in the dye molecules that are prone to rotation. First, when the viscosity of the system is small, the rotation and vibration of the double bond can cause uneven distribution of partial charges, resulting in quenching of fluorescent signals. When the viscosity of the system is increased, the molecules are in a rigid plane, and finally, a brighter fluorescent signal is displayed.
The fluorescent dye provided by the invention constructs the fluorescent dye with large Stokes shift based on fluorescence resonance energy transfer effect (FRET). In the dye, naphthalimide is used as FRET energy donor, and dicyanoisophorone derivative is used as energy acceptor, so that Stokes displacement is increased. By testing the fluorescence emission spectra in various organic solvents, it was found that: stokes displacement is between 173 and 234nm, and has obvious fluorescence signal emission.
Claims (6)
1. A fluorescent dye with large Stokes shift, which is characterized in that the fluorescent dye molecule has a structural formula shown in the following formula (I):
2. a method for preparing a fluorescent dye with a large Stokes shift as claimed in claim 1, which is carried out in particular according to the following steps:
step 1, synthesizing an intermediate TEM (transmission electron microscope) by taking isophorone and malononitrile as raw materials through Knoevenagel condensation reaction;
the structure of intermediate TEM is as follows:
；
step 2, synthesizing an intermediate BMD by taking 4-bromo-1, 8-naphthalene dicarboxylic anhydride and N- (2-aminoethyl) morpholine as raw materials;
the structure of intermediate BMD is as follows:
；
step 3, synthesizing an intermediate NAA by taking the intermediate BMD and 4-piperazine-1-benzaldehyde obtained in the step 2 as raw materials;
the structure of intermediate NAA is shown below:
；
and step 4, synthesizing the fluorescent dye by taking the intermediate TEM obtained in the step 1 and the intermediate NAA obtained in the step 3 as raw materials.
3. The method for preparing a fluorescent dye with large Stokes shift according to claim 2, wherein in the step 1, specifically:
dissolving isophorone, malononitrile and a catalyst in N, N-dimethylformamide, taking argon as a protective gas, stirring for 6 hours at 120 ℃, cooling to room temperature after the reaction is finished, injecting a reaction mixed solution into ice water, separating out brown solid, drying, and separating and purifying by using column chromatography to obtain an intermediate TEM; the catalyst is a viscous liquid formed by mixing acetic anhydride, glacial acetic acid and piperidine; the molar ratio of isophorone to malononitrile is 1:1.
4. The method for preparing a fluorescent dye with large Stokes shift according to claim 2, wherein in the step 2, specifically:
dissolving 4-bromo-1, 8-naphthalene dicarboxylic anhydride and N- (2-aminoethyl) morpholine in 1, 4-dioxane, and reacting at 80 ℃ for 8h; cooling to room temperature after finishing, injecting the reaction solution into ice water to generate precipitate, filtering to obtain light yellow solid, drying, separating and purifying by batch column chromatography, wherein the eluent is dichloromethane to obtain white solid, namely intermediate BMD; the molar ratio of 4-bromo-1, 8-naphthalene dicarboxylic anhydride to N- (2-aminoethyl) morpholine was 1:2.
5. The method for preparing a fluorescent dye with large Stokes shift according to claim 2, wherein in the step 3, specifically:
dissolving BMD, 4-piperazine-1-benzaldehyde and a catalyst in dimethyl sulfoxide, heating and stirring at 100 ℃, tracking by TLC until the reaction is finished, cooling to room temperature, adding water into a reaction liquid to generate yellow solid precipitate, carrying out suction filtration and air drying, and separating and purifying by using column chromatography to finally obtain a yellow-green solid product, namely an intermediate NAA; the molar ratio of BMD to 4-piperazine-1-benzaldehyde is 2.5:3, and cesium carbonate is used as a catalyst.
6. The method for preparing a fluorescent dye with large Stokes shift according to claim 2, wherein in the step 4, specifically:
dissolving an intermediate TEM, an intermediate NAA and a catalyst in absolute ethyl alcohol, taking argon as a protective gas, heating and stirring for 4 hours at 80 ℃, cooling to room temperature after the reaction is finished, filtering to obtain a reddish brown solid product, and naturally airing to obtain the fluorescent dye with large Stokes displacement; the molar ratio of TEM to NAA is 2:3; the catalyst is piperidine.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5279967A (en) * | 1992-01-24 | 1994-01-18 | Nalco Chemical Company | Fluorescent labeling of hydrocarbons for source identification |
| GB2533166A (en) * | 2014-12-12 | 2016-06-15 | Univ Malta | Sensor molecule |
| CN108129459A (en) * | 2018-01-10 | 2018-06-08 | 济南大学 | A kind of novel fluorescence probe and its application for being used to detect sulfur dioxide |
| CN108250220A (en) * | 2018-02-06 | 2018-07-06 | 广西师范学院 | Phenothiazines coumarine dye and synthetic method with big Stokes displacements |
| CN111334078A (en) * | 2018-12-18 | 2020-06-26 | 中国科学院大连化学物理研究所 | 405nm excited high-brightness and high-stability fluorescent dye and synthetic method thereof |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5279967A (en) * | 1992-01-24 | 1994-01-18 | Nalco Chemical Company | Fluorescent labeling of hydrocarbons for source identification |
| GB2533166A (en) * | 2014-12-12 | 2016-06-15 | Univ Malta | Sensor molecule |
| CN108129459A (en) * | 2018-01-10 | 2018-06-08 | 济南大学 | A kind of novel fluorescence probe and its application for being used to detect sulfur dioxide |
| CN108250220A (en) * | 2018-02-06 | 2018-07-06 | 广西师范学院 | Phenothiazines coumarine dye and synthetic method with big Stokes displacements |
| CN111334078A (en) * | 2018-12-18 | 2020-06-26 | 中国科学院大连化学物理研究所 | 405nm excited high-brightness and high-stability fluorescent dye and synthetic method thereof |
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