CN102702768B - Novel red BODIPY fluorescent dye and preparation method and application thereof - Google Patents
Novel red BODIPY fluorescent dye and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to a novel red BODIPY fluorescent dye with the chemical formula of C10+mH7+nBF2N2+xOy, wherein m, n, x and y are integers from 0 to 100. The preparation method comprises the following steps: dissolving pyrrole with substituent groups R1, R2 and R3 in an organic solution; adding ethyl glyoxylate together with nitrogen to the organic solution for a chemical reaction by using trifluoroacetic acid or toluenesulfonic acid as a catalyst; adding 2,3-dichloro-5,6-dicyano-1,4-benzoquinone oxidative dehydrogenation; and adding organic amine and a boron trifluoride diethyl ether solution for another reaction. After the reaction solution is concentrated, chromatography is performed with a silicagel column to obtain the fluorescent dye. The fluorescent dye can be used for cell imaging, fluorescent probe or laser dye. The fluorescent dye has the advantages that the ultraviolet-visible absorption spectrum and the fluorescence emission spectrum of the fluorescent dye are narrow; fluorescent quanta has high efficiency and good light stability; and the fluorescent dye has simple molecular structure and can be synthesized easily, so as to facilitate popularization and application.
Description
Technical field
The present invention relates to fluorescence dye and its preparation method and application, particularly a kind of novel ruddiness BODIPY fluorescence dye and its preparation method and application.
Background technology
Boron fluoride network and two pyrroles's methine fluorescence dyes (BODIPY) are the class novel fluorescence dyestuffs occurred in recent years, and it is the mixture formed by two pyrroles's methylene radical and boron trifluoride.This dye molecule has higher stability, can avoid dyestuff from the change of the rising of the irradiation of stimulated luminescence in the fluorometric analysis process, temperature or testing environment and cause dye structure to be degraded rapidly, has also guaranteed the stability of spectroscopic analysis signal.In addition, the BODIPY dyestuff has advantages of that fluorescence quantum efficiency is high, absorption spectrum and fluorescence emission spectrum narrow, thereby there is very high sensitivity when bioanalysis and biomarker.
1985, the people such as Worries H.J. and Koek J.H. synthesized the BODIPY dyestuff that 2 bit strips have sulfonic acid group, and this dyestuff exists with sodium-salt form, therefore had well water-soluble.This is successfully in BODIPY dye matrix structure, to have introduced active group first, for the BODIPY dyestuff is widely used in the bioanalysis field in the future, has walked out the first step.In order to solve the water-soluble of BODIPY molecule, the people such as Ziessel R. introduce quaternary ammonium salt groups at 2,6 of BODIPY, have obtained a series of good water solubility, BODIPY fluorescence dye that fluorescence quantum efficiency is high.Liu H.Y. passes through to introduce water miscible polyethylene group at 3,5 of BODIPY, and replaces BODIPY center fluorine atom with the alkynes reagent of polyoxyethylene glycol functionalization, obtains the fluorescence dye of some row good water solubility.
In recent years, the BODIPY dyestuff to the research trend of BODIPY dyestuff in synthetic long wavelength, because the emission wavelength of dyestuff is longer, the light penetration power is strong, background interference is little, can be advantageously applied to bioanalysis and bio-imaging.For the fluorescent emission wavelength adjustment BODIPY, to near-infrared region, the researchist has mainly used following three kinds of methods:
1) pyrroles is changed into to isoquinoline 99.9 and the furo pyrroles increases the pi-conjugated structure of molecule, and be connected aromatic substituent at 3 with 5.Nineteen ninety-five, it is raw material that Haugland R.P. and Kang H.C. adopt adjacent benzene dicarbonyl compound, first with azanol reaction, generates isoindole dimethime intermediate, then with the boron trifluoride complexing, finally obtains having the BODIPY fluorescence dye of large conjugated system.The maximum emission wavelength of this serial BODIPY dye matrix structure is less than 600nm, but, by after 3,5 of the parent nucleus thienyls that connect two thienyls or have alkyl to replace, maximum emission wavelength has even surpassed 700nm, becomes near infrared fluorescent dye.But this series of compound, because the solvability that can't introduce attachable group and dye molecule is very poor, is difficult to permeates cell membranes and enters in cell tissue, thereby hindered its application in biochemical field.
2) connect in the α position of BODIPY conjugation the aromatic group that pushes away electronics.The people such as Haugland R.P. and Kang H.C. couple together aromatic group and BODIPY dye matrix by etheno, the fluorescent emission wavelength that obtains dyestuff is 652nm, explanation is introduced styryl on 2 of BODIPY dyestuff parent nucleus, can make the emission wavelength red shift greatly of dyestuff.Burgess study group has synthesized a series of furo azole compounds, and researched and developed as raw material and acyl chlorides condensation the glimmering dyestuff of a class fluorine boron that maximum emission wavelength is greater than 650nm, this class dyestuff has reduced the torsion degree of molecule, thereby makes its maximum emission wavelength red shift.
3) replace 8 upper carbon atoms of common BODIPY with nitrogen-atoms, obtain aza-BODIPY.Synthetic have that large stokes (Stokes) displacement, fluorescence quantum efficiency are high in order to design, good light stability and make the fluorescent emission wavelength of BODIPY molecule arrive near-infrared region, O ' Shea ' study group replaces 8 upper carbon atoms of common BODIPY with nitrogen-atoms, obtain aza-BODIPY.The molar extinction coefficient of aza-BODIPY dyestuff is large, fluorescence quantum yield is high, fluorescence spectrum half-peak width, fluorescence lifetime long and good light stability, and its fluorescence is all insensitive to polarity and the pH of solvent, be the fluorescence dye that a class can be applicable to biological field, very likely as photosensitizers, be applied to photodynamic therapy.
The poorly water-soluble of most BODIPY dyestuff, lacking can be for the group of functionalization, thereby has limited its further application on biomedicine.Better in order to design synthesizing water-solubility, the near infrared fluorescent dye of active function groups is arranged, the present invention designs and has synthesized a kind of novel structure, excellent property, the simple novel B ODIPY fluorescence dye of preparation method.
Summary of the invention
The objective of the invention is to overcome existing fluorescence dye in performance and structural deficiency, provide that a kind of molecular structure is simple, fluorescence quantum efficiency is high, emission wavelength is long, good light stability and the simple novel ruddiness BODIPY fluorescence dye of preparation method, this fluorescence dye is applicable to cell imaging, fluorescent probe or laser dyes.
Technical scheme of the present invention:
A kind of novel ruddiness BODIPY fluorescence dye, chemical formula is C
10+mH
7+nBF
2N
2+xO
y, m wherein, n, x, y is the integer of 0-100, and molecule has following structure:
Wherein: R
1, R
2, R
3For alkyl, aryl, thiazolinyl, alkynyl, ester group, ether, thioether group, cyano group or halogen atom; R
4For alkyl, aryl, ether, hydroxyl or amino; R
5, R
6For alkyl, alkoxyl group, ether, thioether group, alkynyl or fluorine atom;
For negative charge,
For positive charge.
A kind of preparation method of described novel ruddiness BODIPY fluorescence dye, step is as follows:
1) will be with substituent R
1, R
2, R
3The pyrroles be dissolved in organic solvent, add oxoethanoic acid second fat under nitrogen atmosphere, obtain mixed solution, in mixed solution organic solvent with substituent R
1, R
2, R
3Pyrroles's volume ratio be 1-10000:1, oxoethanoic acid second fat with substituent R
1, R
2, R
3Pyrroles's mol ratio be 0.01-0.50:1;
2) under nitrogen protection, catalyst drops is added in reaction solution, in temperature, be to react 3 hours under 0-100 ℃, obtain reaction solution;
3) add 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) in above-mentioned reaction solution, the mol ratio of DDQ and oxoethanoic acid second fat is 0.01-10:1, and oxydehydrogenation 0.5 hour, obtain dehydrogenation liquid;
4) add successively organic amine and boron trifluoride ether solution in above-mentioned dehydrogenation liquid, the mol ratio of organic amine and oxoethanoic acid second fat is 1-300:1, the mol ratio of boron trifluoride ether solution and oxoethanoic acid second fat is 0.1-500:1, continues reaction 1 hour, obtains target solution;
5) underpressure distillation is except after desolventizing, and residue obtains target compound through silica gel column chromatography.
Described is R with substituting group
1, R
2, R
3The pyrroles in substituent R
1, R
2, R
3For alkyl, aryl, thiazolinyl, alkynyl, ester group, ether, thioether group, cyano group or halogen atom and be directly connected on pyrrole ring.
Described organic solvent is methylene dichloride, chloroform, benzene,toluene,xylene, ether, ethyl acetate, methyl-sulphoxide or dioxane.
Described catalyzer is trifluoroacetic acid or tosic acid.
Described organic amine is triethylamine, Trimethylamine 99, diisopropylethylamine, diisopropyl aniline or piperidines.
A kind of application of described novel ruddiness BODIPY fluorescence dye, for cell imaging, fluorescent probe or laser dyes, method is: this luminescent dye molecule can be dissolved in the aqueous buffer solution of physiological saline or hydroxyethyl piperazine ethanesulfonic acid (HEPES); Perhaps this luminescent dye molecule is dissolved in the organic solvents such as methyl alcohol, ethanol, acetonitrile, methyl-sulphoxide, diformamide; Perhaps the mixed solvent of water and above-mentioned organic solvent arbitrary proportion, then be directly used in various application scenarios to the above-mentioned solution that is dissolved with this fluorescence dye.
Advantage of the present invention is: ultraviolet-visible absorption spectroscopy and the fluorescence emission spectrum of this fluorescence dye is narrow, fluorescence quantum efficiency is high, good light stability; Can, for cell imaging, fluorescent probe or laser dyes, show good practical application; Luminescent dye molecule of the present invention is simple in structure, simple synthetic method, makes its very easily actual applying.
The accompanying drawing explanation
The fluorescence dye one dimension hydrogen nuclear magnetic spectrum that Fig. 1 is embodiment 1 preparation, in figure: X-coordinate is chemical shift, ordinate zou is strength of signal.
The fluorescence dye ultraviolet-visible absorption spectroscopy that Fig. 2 is embodiment 1 preparation.
The fluorescence dye fluorescence emission spectrum that Fig. 3 is embodiment 1 preparation.
The fluorescence dye ultraviolet-visible absorption spectroscopy that Fig. 4 is embodiment 6 preparations.
The fluorescence dye fluorescence emission spectrum that Fig. 5 is embodiment 6 preparations.
The laser confocal imaging figure of fluorescence dye in tumour cell that Fig. 6 is embodiment 1 preparation.
The fluorescence dye that Fig. 7 is embodiment 2 preparation as fluorescent probe to tumour cell endoplasmic reticulum image.
Embodiment
Embodiment 1:
A kind of preparation method of novel ruddiness BODIPY fluorescence dye, step is as follows:
1) 0.25 milliliter of 2-N-ethyl pyrrole N-is dissolved in 50 milliliters of toluene, adds 0.10 milliliter of oxoethanoic acid second fat under nitrogen atmosphere, obtain mixed solution, in mixed solution organic solvent with substituent R
1, R
2, R
3Pyrroles's volume ratio be 200:1, oxoethanoic acid second fat with substituent R
1, R
2, R
3Pyrroles's mol ratio be 1:2.4;
2) under nitrogen protection, 5 micro litres of catalyst trifluoroacetic acids are added drop-wise in reaction solution, in temperature, be to react 3 hours under 0 ℃, obtain reaction solution;
3) add the 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) of 245 milligrams in above-mentioned reaction solution, the mol ratio of DDQ and oxoethanoic acid second fat is 1.1:1, and oxydehydrogenation 0.5 hour, obtain dehydrogenation liquid;
4) add successively 0.12 milliliter of triethylamine and 0.16 milliliter of boron trifluoride ether solution in above-mentioned dehydrogenation liquid, the mol ratio of triethylamine and oxoethanoic acid second fat is 1.1:1, the mol ratio of boron trifluoride ether solution and oxoethanoic acid second fat is 1.1:1, continues reaction 1 hour, obtains target solution;
5) underpressure distillation is except after desolventizing, and residue obtains target compound through silica gel column chromatography.
The constitutional features of this fluorescence dye:
1H?NMR(500.13MHz,CDCl
3):δ1.34(m,6H),1.44(m,3H),3.04(q,J=7.6Hz,4H),4.47(q,J=6.9Hz,2H),6.39(d,J=3.2Hz,2H),7.26(d,J=3.2Hz,2H).
13C?NMR(125.77MHz,CDCl
3):δ14.2,22.3,29.7,62.5,118.4,128.8,130.7,133.1,163.8,166.4.EI-MS?m/z=320.15M
+。
The fluorescence dye one dimension hydrogen nuclear magnetic spectrum that Fig. 1 is embodiment 1 preparation, in figure: X-coordinate is chemical shift, ordinate zou is strength of signal.In figure, show: each chemical shift of organizing hydrogen is fully corresponding with the structure of this dyestuff, can be confirmed to be our synthetic target compound.
The fluorescence dye ultraviolet-visible absorption spectroscopy that Fig. 2 is embodiment 1 preparation.The fluorescence dye fluorescence emission spectrum that Fig. 3 is embodiment 1 preparation.In figure, show: the ultraviolet maximum absorption wavelength of fluorescence dye is that 525nm(is shown in Fig. 2), the fluorescence maximum emission wavelength is 586nm, and the Stokes displacement is 61nm, and fluorescence quantum efficiency is 0.83, illustrates that this fluorescence dye has good fluorescence property.
Cell cultures and laser confocal imaging experiment:
Get the 1.0x10 that 0.1mL configures
-5The dichloromethane solution of embodiment 1 compound of mol/L, in the volumetric flask of 10mL, after drying up solvent with nitrogen, after the methyl-sulphoxide dissolving with 0.1mL, adds PBS solution (pH=7.4) to be settled to 10mL stand-by.Conventional DMEM nutrient solution for uterus carcinoma HeLa tumour cell (10% foetal calf serum, 0.2% NaHCO
3The aqueous solution, penicillin 100U/mL) the adjustment cell concn is 1.0x10
5Individual/L, be placed in the culture dish of sterility cover slide, in 37 ° of C, 5%CO
2Incubator is cultivated.Clean 3 times with PBS solution again, remove and there is no adherent cell, obtain adherent uterus carcinoma HeLa tumour cell for experiment.Embodiment 1 compound is shown in Fig. 6 to the laser confocal imaging figure of uterus carcinoma HeLa tumour cell.As seen from the figure, after compound dyeing, at 37 ° of C, 5%CO
2The cell that incubator is cultivated 15 minutes presents very strong fluorescence, and cell is high-visible.Illustrate that this fluorescence dye has good imaging effect to uterus carcinoma HeLa tumour cell.
Embodiment 2:
A kind of preparation method of novel ruddiness BODIPY fluorescence dye, step is as follows:
1) 10 milliliters of 2-N-ethyl pyrrole N-s are dissolved in 10 milliliters of methylene dichloride, add 0.10 milliliter of oxoethanoic acid second fat under nitrogen atmospheres, obtain mixed solution, in mixed solution organic solvent with substituent R
1, R
2, R
3Pyrroles's volume ratio be 1:1, oxoethanoic acid second fat with substituent R
1, R
2, R
3Pyrroles's mol ratio be 1:100;
2) under nitrogen protection, 5 micro litres of catalyst tosic acid are added drop-wise in reaction solution, in temperature, be to react 3 hours under 100 ℃, obtain reaction solution;
3) add the 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) of 2.24 grams in above-mentioned reaction solution, the mol ratio of DDQ and oxoethanoic acid second fat is 10.0:1, and oxydehydrogenation 0.5 hour, obtain dehydrogenation liquid;
4) add successively 3 milliliters of Trimethylamine 99s and 4 milliliters of boron trifluoride ether solutions in above-mentioned dehydrogenation liquid, the mol ratio of Trimethylamine 99 and oxoethanoic acid second fat is 30:1, the mol ratio of boron trifluoride ether solution and oxoethanoic acid second fat is 24:1, continues reaction 1 hour, obtains target solution;
5) underpressure distillation is except after desolventizing, and residue obtains target compound through silica gel column chromatography.
The constitutional features of this fluorescence dye:
1H NMR (500.13MHz, CDCl
3): δ 1.34 (m, 6H), 1.44 (m, 3H), 3.04 (q, J=7.6Hz, 4H), 4.47 (q, J=6.9Hz, 2H), 6.39 (d, J=3.2Hz, 2H), 7.26 (d, J=3.2Hz, 2H).
13C NMR (125.77MHz, CDCl
3): δ 14.2,22.3,29.7,62.5,118.4,128.8,130.7,133.1,163.8,166.4.EI-MS m/z=320.15M
+.
Fluorescent probe is at intracellular imaging experiment:
Get the 1.0x10 that 0.1mL configures
-5The dichloromethane solution of embodiment 1 fluorescence dye of mol/L, in the volumetric flask of 10mL, after drying up solvent with nitrogen, after the methyl-sulphoxide dissolving with 0.5mL, adds PBS solution (pH=7.4) to be settled to 10mL stand-by.Conventional DMEM nutrient solution for uterus carcinoma HeLa tumour cell (10% foetal calf serum, 0.2% NaHCO
3The aqueous solution, penicillin 100U/mL) the adjustment cell concn is 1.0x10
5Individual/L, be placed in the culture dish of sterility cover slide, in 37 ° of C, 5%CO
2Incubator is cultivated.Clean 3 times with PBS solution again, remove and there is no adherent cell, obtain adherent uterus carcinoma HeLa tumour cell for experiment.Embodiment 2 compounds are shown in Fig. 7 to the laser confocal imaging figure of uterus carcinoma HeLa tumour cell.As seen from the figure, dyed after, at 37 ° of C, 5%CO
2The endocytoplasmic reticulum that incubator is cultivated 15 minutes presents very strong fluorescence, and endocytoplasmic reticulum is high-visible.Illustrate that this fluorescence dye has the specificity effect to endocytoplasmic reticulum, can be used in the imaging of endocytoplasmic reticulum.
Embodiment 3:
A kind of preparation method of novel ruddiness BODIPY fluorescence dye, step is as follows:
1) 0.2 milliliter of 2-N-ethyl pyrrole N-is dissolved in 50 milliliters of ether, adds 0.10 milliliter of oxoethanoic acid second fat under nitrogen atmosphere, obtain mixed solution, in mixed solution organic solvent with substituent R
1, R
2, R
3Pyrroles's volume ratio be 1:250, oxoethanoic acid second fat with substituent R
1, R
2, R
3Pyrroles's mol ratio be 1:500;
2) under nitrogen protection, 5 micro litres of catalyst trifluoroacetic acids are added drop-wise in reaction solution, in temperature, be to react 3 hours under 20 ℃, obtain reaction solution;
3) add the 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) of 2.3 milligrams in above-mentioned reaction solution, the mol ratio of DDQ and oxoethanoic acid second fat is 0.01:1, and oxydehydrogenation 0.5 hour, obtain dehydrogenation liquid;
4) add successively 30 milliliters of diisopropylethylamine and 80 milliliters of boron trifluoride ether solutions in above-mentioned dehydrogenation liquid, the mol ratio of diisopropylethylamine and oxoethanoic acid second fat is 30:1, the mol ratio of boron trifluoride ether solution and oxoethanoic acid second fat is 50:1, continue reaction 1 hour, obtain target solution;
5) underpressure distillation is except after desolventizing, and residue obtains target compound through silica gel column chromatography.
The constitutional features of this fluorescence dye:
1H NMR (500.13MHz, CDCl
3): δ 1.34 (m, 6H), 1.44 (m, 3H), 3.04 (q, J=7.6Hz, 4H), 4.47 (q, J=6.9Hz, 2H), 6.39 (d, J=3.2Hz, 2H), 7.26 (d, J=3.2Hz, 2H).
13C NMR (125.77MHz, CDCl
3): δ 14.2,22.3,29.7,62.5,118.4,128.8,130.7,133.1,163.8,166.4.EI-MS m/z=320.15M
+.
Embodiment 4:
A kind of preparation method of novel ruddiness BODIPY fluorescence dye, step is as follows:
1) 0.2 milliliter of 2-N-ethyl pyrrole N-is dissolved in 50 milliliters of chloroforms, adds 0.10 milliliter of oxoethanoic acid second fat under nitrogen atmosphere, obtain mixed solution, in mixed solution organic solvent with substituent R
1, R
2, R
3Pyrroles's volume ratio be 1:250, oxoethanoic acid second fat with substituent R
1, R
2, R
3Pyrroles's mol ratio be 1:500;
2) under nitrogen protection, 5 micro litres of catalyst tosic acid are added drop-wise in reaction solution, in temperature, be to react 3 hours under 20 ℃, obtain reaction solution;
3) add the 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) of 245 milligrams in above-mentioned reaction solution, the mol ratio of DDQ and oxoethanoic acid second fat is 1.1:1, and oxydehydrogenation 0.5 hour, obtain dehydrogenation liquid;
4) add successively 0.1 milliliter of diisopropyl aniline and 0.02 milliliter of boron trifluoride ether solution in above-mentioned dehydrogenation liquid, the mol ratio of diisopropyl aniline and oxoethanoic acid second fat is 1.1:1, the mol ratio of boron trifluoride ether solution and oxoethanoic acid second fat is 0.1:1, continue reaction 1 hour, obtain target solution;
5) underpressure distillation is except after desolventizing, and residue obtains target compound through silica gel column chromatography.
The constitutional features of this fluorescence dye:
1H NMR (500.13MHz, CDCl
3): δ 1.34 (m, 6H), 1.44 (m, 3H), 3.04 (q, J=7.6Hz, 4H), 4.47 (q, J=6.9Hz, 2H), 6.39 (d, J=3.2Hz, 2H), 7.26 (d, J=3.2Hz, 2H).
13C NMR (125.77MHz, CDCl
3): δ 14.2,22.3,29.7,62.5,118.4,128.8,130.7,133.1,163.8,166.4.EIMS m/z=320.15M
+.
Embodiment 5:
A kind of preparation method of novel ruddiness BODIPY fluorescence dye, step is as follows:
1) by 0.25 milliliter 2, the 4-dimethyl pyrrole is dissolved in 50 milliliters of methylene dichloride, adds 0.10 milliliter of oxoethanoic acid second fat under nitrogen atmosphere, obtains mixed solution, in mixed solution organic solvent with substituent R
1, R
2, R
3Pyrroles's volume ratio be 200:1, oxoethanoic acid second fat with substituent R
1, R
2, R
3Pyrroles's mol ratio be 1:2.4;
2) under nitrogen protection, 5 micro litres of catalyst trifluoroacetic acids are added drop-wise in reaction solution, in temperature, be to react 3 hours under 20 ℃, obtain reaction solution;
3) add the 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) of 245 milligrams in above-mentioned reaction solution, the mol ratio of DDQ and oxoethanoic acid second fat is 1.1:1, and oxydehydrogenation 0.5 hour, obtain dehydrogenation liquid;
4) add successively 0.12 milliliter of piperidines and 0.16 milliliter of boron trifluoride ether solution in above-mentioned dehydrogenation liquid, the mol ratio of piperidines and oxoethanoic acid second fat is 1.1:1, the mol ratio of boron trifluoride ether solution and oxoethanoic acid second fat is 1.1:1, continues reaction 1 hour, obtains target solution;
5) underpressure distillation is except after desolventizing, and residue obtains target compound through silica gel column chromatography.
The constitutional features of this fluorescence dye:
1H NMR (500.13MHz, CDCl
3): δ 1.38 (s, 6H), 1.44 (m, 3H), 2.09 (s, 6H), 4.47 (q, J=6.9Hz, 2H), 6.02 (s, 2H).
13C NMR (125.77MHz, CDCl
3): δ 22.3,25.9,29.7,62.5,118.48,128.8,130.2,133.7,163.5,166.1.EI-MS m/z=320.15M
+.
Embodiment 6:
A kind of preparation method of novel ruddiness BODIPY fluorescence dye, step is as follows:
1) by 0.25 milliliter 2, the 4-dimethyl pyrrole is dissolved in 50 milliliters of dimethylbenzene, adds 0.10 milliliter of oxoethanoic acid second fat under nitrogen atmosphere, obtains mixed solution, in mixed solution organic solvent with substituent R
1, R
2, R
3Pyrroles's volume ratio be 200:1, oxoethanoic acid second fat with substituent R
1, R
2, R
3Pyrroles's mol ratio be 1:2.4;
2) under nitrogen protection, 5 micro litres of catalyst trifluoroacetic acids are added drop-wise in reaction solution, in temperature, be to react 3 hours under 20 ℃, obtain reaction solution;
3) add the 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) of 245 milligrams in above-mentioned reaction solution, the mol ratio of DDQ and oxoethanoic acid second fat is 1.1:1, and oxydehydrogenation 0.5 hour, obtain dehydrogenation liquid;
4) add successively 0.12 milliliter of triethylamine and 0.16 milliliter of boron trifluoride ether solution in above-mentioned dehydrogenation liquid, the mol ratio of triethylamine and oxoethanoic acid second fat is 1.1:1, the mol ratio of boron trifluoride ether solution and oxoethanoic acid second fat is 1.1:1, continues reaction 1 hour, obtains target solution;
5) underpressure distillation is except after desolventizing, and residue obtains target compound through silica gel column chromatography.
The constitutional features of this fluorescence dye:
1H NMR (500.13MHz, CDCl
3): δ 1.38 (s, 6H), 1.44 (m, 3H), 2.09 (s, 6H), 4.47 (q, J=6.9Hz, 2H), 6.02 (s, 2H).
13C NMR (125.77MHz, CDCl
3): δ 22.3,25.9,29.7,62.5,118.48,128.8,130.2,133.7,163.5,166.1.EI-MS m/z=320.15M
+.
Fig. 5 is that (concentration is 1.0x10 to this fluorescence dye
-7Mol/L) fluorescence emission spectrum in methylene dichloride.The maximum absorption wavelength of fluorescence dye is that 596nm(is shown in Fig. 4), the fluorescence maximum emission wavelength is 676nm, and the Stokes displacement is 80nm, and fluorescence quantum efficiency is 0.23, illustrates that this fluorescence dye has good fluorescence property.
Claims (4)
1. the preparation method of a novel ruddiness BODIPY fluorescence dye, the chemical formula of described fluorescence dye is C
10+mH
7+nBF
2N
2+xO
y, m wherein, n, x is the integer of 0-100, the integer that y is 2-100, and molecule has following structure:
Wherein: R
1, R
2, R
3For alkyl, aryl, thiazolinyl, alkynyl, ester group or ether; R
4For alkoxyl oxygen alkyl ethyl, R
5, R
6For fluorine atom,
For negative charge, ⊕ is positive charge, it is characterized in that step is as follows:
1) will be with substituent R
1, R
2, R
3The pyrroles be dissolved in organic solvent, add oxoethanoic acid second fat under nitrogen atmosphere, obtain mixed solution, in mixed solution organic solvent with substituent R
1, R
2, R
3Pyrroles's volume ratio be 1-10000:1, oxoethanoic acid second fat with substituent R
1, R
2, R
3Pyrroles's mol ratio be 0.01-0.50:1;
2) under nitrogen protection, catalyst drops is added in reaction solution, in temperature, be to react 3 hours under 0-100 ℃, obtain reaction solution;
3) add 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) in above-mentioned reaction solution, the mol ratio of DDQ and oxoethanoic acid second fat is 0.01-10.0:1, and oxydehydrogenation 0.5 hour, obtain dehydrogenation liquid;
4) add successively organic amine and boron trifluoride ether solution in above-mentioned dehydrogenation liquid, the mol ratio of organic amine and oxoethanoic acid second fat is 1.1-300:1, the mol ratio of boron trifluoride ether solution and oxoethanoic acid second fat is 0.1-500:1, continues reaction 1 hour, obtains target solution;
5) underpressure distillation is except after desolventizing, and residue obtains target compound through silica gel column chromatography.
2. the preparation method of novel ruddiness BODIPY fluorescence dye according to claim 1, it is characterized in that: described organic solvent is methylene dichloride, chloroform, benzene,toluene,xylene, ether, tetrahydrofuran (THF), ethyl acetate, methyl-sulphoxide or dioxane.
3. the preparation method of novel ruddiness BODIPY fluorescence dye according to claim 1, it is characterized in that: described catalyzer is trifluoroacetic acid or tosic acid.
4. the preparation method of novel ruddiness BODIPY fluorescence dye according to claim 1, it is characterized in that: described organic amine is triethylamine, Trimethylamine 99, diisopropylethylamine, diisopropyl aniline or piperidines.
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| CN102993763B (en) * | 2012-12-09 | 2014-08-27 | 大连理工大学 | Single charge boron fluroride complexing dipyrrole methenyl fluorochrome and application thereof |
| CN103242823B (en) * | 2013-04-28 | 2014-11-12 | 孔凯明 | O,O two-tooth type organic boron difluoride fluorescent dye and preparation method thereof |
| CN103666456B (en) * | 2013-12-02 | 2015-02-04 | 大连理工大学 | Boron-dipyrromethene fluorescence probes, and making method and application thereof |
| CN104073018B (en) * | 2014-05-09 | 2016-09-28 | 天津大学 | A series of 2,3,5,6-tetra-thiophene with longwave absorption characteristic replaces BODIPY dyestuff and preparation method |
| CN104356680B (en) * | 2014-11-18 | 2017-01-04 | 沈阳化工大学 | Water solublity PnO2-PODIPY/PnO2-azaPODIPY fluorescent dye and preparation method thereof |
| CN104403663B (en) * | 2014-12-11 | 2016-09-28 | 华东理工大学 | A kind of detection endogenous H2fluorescent probe of S and its preparation method and application |
| CN104650610A (en) * | 2015-01-28 | 2015-05-27 | 天津理工大学 | Asymmetric near-infrared BODIPY fluorescent dye as well as preparation method and application thereof |
| CN105175310B (en) * | 2015-05-22 | 2020-11-13 | 南京大学 | Asymmetric aza diisoindole methylene compound and its synthesis and use of boron fluoride compound |
| CN111333670B (en) * | 2019-12-12 | 2023-05-16 | 安徽师范大学 | Isoindole fluorine boron tripyrrole fluorescent dye and preparation method thereof |
| CN111100154B (en) * | 2019-12-31 | 2022-02-22 | 华侨大学 | Benzoic acid substituted BODIPY derivative dye ligand and preparation method thereof |
| CN113912632B (en) * | 2021-11-10 | 2023-11-17 | 四川大学华西医院 | Fluoroboron fluorescent dye and preparation method and application thereof |
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