CN102702768A - 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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- CN102702768A CN102702768A CN2012101808100A CN201210180810A CN102702768A CN 102702768 A CN102702768 A CN 102702768A CN 2012101808100 A CN2012101808100 A CN 2012101808100A CN 201210180810 A CN201210180810 A CN 201210180810A CN 102702768 A CN102702768 A CN 102702768A
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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 optical dye, particularly a kind of novel red light BODIPY optical dye.
Background technology
Boron fluoride network and two pyrroles's methine optical dyes (BODIPY) are the one type of novel fluorescence dyestuffs that occurs in recent years, and it is the mixture that is formed by two pyrroles's methylene radical and boron trifluoride.This dye molecule has advantages of 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 the narrow advantage of fluorescence quantum efficiency height, absorption spectrum and fluorescence emission spectrum, thereby when bioanalysis and biomarker, has very high sensitivity.
1985, 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 reactive group first, has walked out the first step for the BODIPY dyestuff is widely used in the bioanalysis field in the future.In order to solve the water-soluble of BODIPY molecule, people such as Ziessel R. introduce quaternary ammonium salt groups 2,6 of BODIPY, have obtained a series of good water solubility, BODIPY optical 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 optical dye of some row good water solubility.
In recent years, the research of BODIPY dyestuff is tended to synthesize long wavelength's BODIPY dyestuff, because the emission wavelength of dyestuff is longer, the light penetration power is strong, and background interference is little, can be advantageously applied to bioanalysis and bio-imaging.In order to arrive the near-infrared region to the fluorescent emission wavelength adjustment of BODIPY, the researchist has mainly used following three kinds of methods:
1) changes the pyrroles into pi-conjugated structure that isoquinoline 99.9 and furo pyrroles increase molecule, and be connected aromatic substituent with 5 at 3.Nineteen ninety-five, it is raw material that Haugland R.P. and Kang H.C. adopt adjacent benzene dicarbonyl compound, generates isoindole dimethime midbody with azanol reaction earlier, then with the boron trifluoride complexing, finally obtains having the BODIPY optical dye of big conjugated system.The maximum emission wavelength of this serial BODIPY dye matrix structure is less than 600nm, but after 3,5 of parent nucleus are connected two thienyls or the substituted thienyl of alkyl is arranged, and maximum emission wavelength even surpassed 700nm becomes near infrared fluorescent dye.But this a series of compound is difficult to permeates cell membranes and gets in the cell tissue, thereby hindered its application in biochemical field owing to the solvability that can't introduce attachable group and dye molecule is very poor.
2) conjugation connects the aromatic group that pushes away electronics in the α position of BODIPY.People such as Haugland R.P. and Kang H.C. couple together aromatic group and BODIPY dye matrix through 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 cpds; And researched and developed the one type fluorine boron glimmering dyestuff of maximum emission wavelength greater than 650nm as raw material and acyl chlorides condensation; This type dyestuff has reduced the degree of reversing of molecule, thereby makes its maximum emission wavelength red shift.
3) replace 8 of common BODIPY with nitrogen-atoms and go up carbon atoms, obtain aza-BODIPY.In order to design the synthetic fluorescent emission wavelength arrival near-infrared region that has big stokes (Stokes) displacement, fluorescence quantum efficiency height, good light stability and make the BODIPY molecule; O ' Shea ' study group obtains aza-BODIPY with 8 last carbon atoms of the common BODIPY of nitrogen-atoms replacement.The molar extinction coefficient of aza-BODIPY dyestuff is big, 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 of solvent and pH; Be one type of optical dye that can be applicable to biological field, very likely be applied to photodynamic therapy as photosensitizers.
The poorly water-soluble of present most of BODIPY dyestuffs lacks the group that can be used for 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 optical dye of preparation method.
Summary of the invention
The objective of the invention is to overcome existing optical 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 red light BODIPY optical dye of preparation method, this optical dye is applicable to cell imaging, fluorescent probe or laser dyes.
Technical scheme of the present invention:
A kind of novel red light BODIPY optical dye, chemical formula are C
10+mH
7+nBF
2N
2+xO
y, m wherein, n, x, y are the integer of 0-100, and molecule has following structure:
Wherein: R
1, R
2, R
3Be alkyl, aryl, thiazolinyl, alkynyl, ester group, ether, thioether group, cyanic acid or halogen atom; R
4Be alkyl, aryl, ether, hydroxyl or amino; R
5, R
6Be alkyl, alkoxyl group, ether, thioether group, alkynyl or fluorine atom;
Be negative charge,
Be positive charge.
A kind of preparation method of said novel red light BODIPY optical dye, step is following:
1) will have substituent R
1, R
2, R
3The pyrroles be dissolved in the organic solvent, under nitrogen atmosphere, add oxoethanoic acid second fat, obtain mixed solution, in the mixed solution organic solvent with have substituent R
1, R
2, R
3Pyrroles's volume ratio be 1-10000:1, oxoethanoic acid second fat with have substituent R
1, R
2, R
3Pyrroles's mol ratio be 0.01-0.50:1;
2) under nitrogen protection, be added to catalyst drops in the reaction solution, be 0-100 ℃ in temperature and reacted 3 hours down, obtain reaction solution;
3) in above-mentioned reaction solution, add 2,3-two chloro-5,6-dicyano-1,4-benzoquinones (DDQ), the mol ratio of DDQ and oxoethanoic acid second fat is 0.01-10:1, oxydehydrogenation 0.5 hour obtains dehydrogenation liquid;
4) in above-mentioned dehydrogenation liquid, add organic amine and boron trifluoride ether solution successively; 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) after underpressure distillation removed and desolvates, residue obtained target compound through silica gel column chromatography.
The said substituting group that has is R
1, R
2, R
3The pyrroles in substituent R
1, R
2, R
3For alkyl, aryl, thiazolinyl, alkynyl, ester group, ether, thioether group, cyanic acid or halogen atom and be directly connected on the pyrrole ring.
Said organic solvent is methylene dichloride, chloroform, benzene,toluene,xylene, ether, ethyl acetate, methyl-sulphoxide or dioxane.
Said catalyzer is trifluoroacetic acid or tosic acid.
Said organic amine is triethylamine, Trimethylamine 99, diisopropylethylamine, diisopropyl aniline or piperidines.
A kind of application of said novel red light BODIPY optical dye is used for cell imaging, fluorescent probe or laser dyes, and method is: can this luminescent dye molecule be dissolved in the aqueous buffer solution of saline water or HEPES (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 directly is used for various application scenarios to the above-mentioned solution that is dissolved with this optical dye then.
Advantage of the present invention is: the ultraviolet-visible absorption spectroscopy and the fluorescence emission spectrum of this optical dye is narrow, fluorescence quantum efficiency is high, good light stability; Can be used for cell imaging, fluorescent probe or laser dyes, show favorable actual application property; Simple in structure, the simple synthetic method of luminescent dye molecule of the present invention makes its very easily actual applying.
Description of drawings
Fig. 1 is the optical dye one dimension proton magnetic chart spectrum of embodiment 1 preparation, and among the figure: X-coordinate is chemical shift, and ordinate zou is a strength of signal.
Fig. 2 is the optical dye ultraviolet-visible absorption spectroscopy of embodiment 1 preparation.
Fig. 3 is the optical dye fluorescence emission spectrum of embodiment 1 preparation.
Fig. 4 is the optical dye ultraviolet-visible absorption spectroscopy of embodiment 6 preparations.
Fig. 5 is the optical dye fluorescence emission spectrum of embodiment 6 preparations.
Fig. 6 is the laser co-focusing image of optical dye in tumour cell of embodiment 1 preparation.
Fig. 7 be the optical dye of embodiment 2 preparation as fluorescent probe to tumour cell endoplasmic reticulum image.
Embodiment
Embodiment 1:
A kind of preparation method of novel red light BODIPY optical dye, step is following:
1) 0.25 milliliter of 2-N-ethyl pyrrole N-is dissolved in 50 milliliters of toluene, under nitrogen atmosphere, adds 0.10 milliliter of oxoethanoic acid second fat, obtain mixed solution, in the mixed solution organic solvent with have substituent R
1, R
2, R
3Pyrroles's volume ratio be 200:1, oxoethanoic acid second fat with have substituent R
1, R
2, R
3Pyrroles's mol ratio be 1:2.4;
2) under nitrogen protection, be added drop-wise to 5 micro litres of catalyst trifluoroacetic acids in the reaction solution, be 0 ℃ in temperature and reacted 3 hours down, obtain reaction solution;
3) in above-mentioned reaction solution, add 245 milligrams 2,3-two chloro-5,6-dicyano-1,4-benzoquinones (DDQ), the mol ratio of DDQ and oxoethanoic acid second fat is 1.1:1, oxydehydrogenation 0.5 hour obtains dehydrogenation liquid;
4) in above-mentioned dehydrogenation liquid, add 0.12 milliliter of triethylamine and 0.16 milliliter of boron trifluoride ether solution successively; 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) after underpressure distillation removed and desolvates, residue obtained target compound through silica gel column chromatography.
The constitutional features of this optical 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
+。
Fig. 1 is the optical dye one dimension proton magnetic chart spectrum of embodiment 1 preparation, and among the figure: X-coordinate is chemical shift, and ordinate zou is a strength of signal.Show among the figure: it is corresponding fully with the structure of this dyestuff that each organizes the chemical shift of hydrogen, can confirm it is our synthetic target compound.
Fig. 2 is the optical dye ultraviolet-visible absorption spectroscopy of embodiment 1 preparation.Fig. 3 is the optical dye fluorescence emission spectrum of embodiment 1 preparation.Show among the figure: the ultraviolet maximum absorption wavelength of optical dye is the 525nm (see figure 2), and the fluorescence maximum emission wavelength is 586nm, and the Stokes displacement is 61nm, and fluorescence quantum efficiency is 0.83, explains that this optical dye has good fluorescence property.
Cell cultures and laser co-focusing 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, dry up solvent with nitrogen after, after the methyl-sulphoxide dissolving with 0.1mL, adding PBS solution (pH=7.4), to be settled to 10mL for use.Uterus carcinoma HeLa tumour cell is with conventional DMEM nutrient solution (10% foetal calf serum, 0.2% NaHCO
3The aqueous solution, penicillium mould 100U/mL) the adjustment cell concn is 1.0x10
5Individual/L, place the petridish of sterility cover slide, in 37 ° of C, 5%CO
2Incubator is cultivated.Clean 3 times with PBS solution, removing does not have adherent cell again, obtains adherent uterus carcinoma HeLa tumour cell and supplies experiment to use.Embodiment 1 compound is seen Fig. 6 to the laser co-focusing image of uterus carcinoma HeLa tumour cell.Visible by figure, after compound dyeing, at 37 ° of C, 5%CO
2The cell that incubator was cultivated 15 minutes presents very strong fluorescence, and cell is high-visible.Explain that this optical dye has good imaging effect to uterus carcinoma HeLa tumour cell.
Embodiment 2:
A kind of preparation method of novel red light BODIPY optical dye, step is following:
1) 10 milliliters of 2-N-ethyl pyrrole N-s are dissolved in 10 milliliters of methylene dichloride, under nitrogen atmosphere, add 0.10 milliliter of oxoethanoic acid second fat, obtain mixed solution, in the mixed solution organic solvent with have substituent R
1, R
2, R
3Pyrroles's volume ratio be 1:1, oxoethanoic acid second fat with have substituent R
1, R
2, R
3Pyrroles's mol ratio be 1:100;
2) under nitrogen protection, be added drop-wise to 5 micro litres of catalyst tosic acid in the reaction solution, be 100 ℃ in temperature and reacted 3 hours down, obtain reaction solution;
3) in above-mentioned reaction solution, add 2 of 2.24 grams, 3-two chloro-5,6-dicyano-1,4-benzoquinones (DDQ), the mol ratio of DDQ and oxoethanoic acid second fat is 10.0:1, oxydehydrogenation 0.5 hour obtains dehydrogenation liquid;
4) in above-mentioned dehydrogenation liquid, add 3 milliliters of Trimethylamine 99s and 4 milliliters of boron trifluoride ether solutions successively; 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) after underpressure distillation removed and desolvates, residue obtained target compound through silica gel column chromatography.
The constitutional features of this optical 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 optical dye of mol/L in the volumetric flask of 10mL, dry up solvent with nitrogen after, after the methyl-sulphoxide dissolving with 0.5mL, adding PBS solution (pH=7.4), to be settled to 10mL for use.Uterus carcinoma HeLa tumour cell is with conventional DMEM nutrient solution (10% foetal calf serum, 0.2% NaHCO
3The aqueous solution, penicillium mould 100U/mL) the adjustment cell concn is 1.0x10
5Individual/L, place the petridish of sterility cover slide, in 37 ° of C, 5%CO
2Incubator is cultivated.Clean 3 times with PBS solution, removing does not have adherent cell again, obtains adherent uterus carcinoma HeLa tumour cell and supplies experiment to use.Embodiment 2 compounds are seen Fig. 7 to the laser co-focusing image of uterus carcinoma HeLa tumour cell.Visible by figure, dyed after, at 37 ° of C, 5%CO
2The endocytoplasmic reticulum that incubator was cultivated 15 minutes presents very strong fluorescence, and endocytoplasmic reticulum is high-visible.Explain that this optical dye pair cell endoplasmic reticulum has the specificity effect, can be used in the imaging of endocytoplasmic reticulum.
Embodiment 3:
A kind of preparation method of novel red light BODIPY optical dye, step is following:
1) 0.2 milliliter of 2-N-ethyl pyrrole N-is dissolved in 50 milliliters of ether, under nitrogen atmosphere, adds 0.10 milliliter of oxoethanoic acid second fat, obtain mixed solution, in the mixed solution organic solvent with have substituent R
1, R
2, R
3Pyrroles's volume ratio be 1:250, oxoethanoic acid second fat with have substituent R
1, R
2, R
3Pyrroles's mol ratio be 1:500;
2) under nitrogen protection, be added drop-wise to 5 micro litres of catalyst trifluoroacetic acids in the reaction solution, be 20 ℃ in temperature and reacted 3 hours down, obtain reaction solution;
3) in above-mentioned reaction solution, add 2.3 milligrams 2,3-two chloro-5,6-dicyano-1,4-benzoquinones (DDQ), the mol ratio of DDQ and oxoethanoic acid second fat is 0.01:1, oxydehydrogenation 0.5 hour obtains dehydrogenation liquid;
4) in above-mentioned dehydrogenation liquid, add 30 milliliters of diisopropylethylamine and 80 milliliters of boron trifluoride ether solutions successively; 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) after underpressure distillation removed and desolvates, residue obtained target compound through silica gel column chromatography.
The constitutional features of this optical 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 red light BODIPY optical dye, step is following:
1) 0.2 milliliter of 2-N-ethyl pyrrole N-is dissolved in 50 milliliters of chloroforms, under nitrogen atmosphere, adds 0.10 milliliter of oxoethanoic acid second fat, obtain mixed solution, in the mixed solution organic solvent with have substituent R
1, R
2, R
3Pyrroles's volume ratio be 1:250, oxoethanoic acid second fat with have substituent R
1, R
2, R
3Pyrroles's mol ratio be 1:500;
2) under nitrogen protection, be added drop-wise to 5 micro litres of catalyst tosic acid in the reaction solution, be 20 ℃ in temperature and reacted 3 hours down, obtain reaction solution;
3) in above-mentioned reaction solution, add 245 milligrams 2,3-two chloro-5,6-dicyano-1,4-benzoquinones (DDQ), the mol ratio of DDQ and oxoethanoic acid second fat is 1.1:1, oxydehydrogenation 0.5 hour obtains dehydrogenation liquid;
4) in above-mentioned dehydrogenation liquid, add 0.1 milliliter of diisopropyl aniline and 0.02 milliliter of boron trifluoride ether solution successively; 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) after underpressure distillation removed and desolvates, residue obtained target compound through silica gel column chromatography.
The constitutional features of this optical 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 red light BODIPY optical dye, step is following:
1) with 0.25 milliliter 2, the 4-dimethyl pyrrole is dissolved in 50 milliliters of methylene dichloride, under nitrogen atmosphere, adds 0.10 milliliter of oxoethanoic acid second fat, obtains mixed solution, in the mixed solution organic solvent with have substituent R
1, R
2, R
3Pyrroles's volume ratio be 200:1, oxoethanoic acid second fat with have substituent R
1, R
2, R
3Pyrroles's mol ratio be 1:2.4;
2) under nitrogen protection, be added drop-wise to 5 micro litres of catalyst trifluoroacetic acids in the reaction solution, be 20 ℃ in temperature and reacted 3 hours down, obtain reaction solution;
3) in above-mentioned reaction solution, add 245 milligrams 2,3-two chloro-5,6-dicyano-1,4-benzoquinones (DDQ), the mol ratio of DDQ and oxoethanoic acid second fat is 1.1:1, oxydehydrogenation 0.5 hour obtains dehydrogenation liquid;
4) in above-mentioned dehydrogenation liquid, add 0.12 milliliter of piperidines and 0.16 milliliter of boron trifluoride ether solution successively; 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) after underpressure distillation removed and desolvates, residue obtained target compound through silica gel column chromatography.
The constitutional features of this optical 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 red light BODIPY optical dye, step is following:
1) with 0.25 milliliter 2, the 4-dimethyl pyrrole is dissolved in 50 milliliters of YLENE, under nitrogen atmosphere, adds 0.10 milliliter of oxoethanoic acid second fat, obtains mixed solution, in the mixed solution organic solvent with have substituent R
1, R
2, R
3Pyrroles's volume ratio be 200:1, oxoethanoic acid second fat with have substituent R
1, R
2, R
3Pyrroles's mol ratio be 1:2.4;
2) under nitrogen protection, be added drop-wise to 5 micro litres of catalyst trifluoroacetic acids in the reaction solution, be 20 ℃ in temperature and reacted 3 hours down, obtain reaction solution;
3) in above-mentioned reaction solution, add 245 milligrams 2,3-two chloro-5,6-dicyano-1,4-benzoquinones (DDQ), the mol ratio of DDQ and oxoethanoic acid second fat is 1.1:1, oxydehydrogenation 0.5 hour obtains dehydrogenation liquid;
4) in above-mentioned dehydrogenation liquid, add 0.12 milliliter of triethylamine and 0.16 milliliter of boron trifluoride ether solution successively; 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) after underpressure distillation removed and desolvates, residue obtained target compound through silica gel column chromatography.
The constitutional features of this optical 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 optical dye
-7Mol) fluorescence emission spectrum in methylene dichloride.The maximum absorption wavelength of optical dye is the 596nm (see figure 4), and the fluorescence maximum emission wavelength is 676nm, and the Stokes displacement is 80nm, and fluorescence quantum efficiency is 0.23, explains that this optical dye has good fluorescence property.
Claims (7)
1. novel red light BODIPY optical dye, it is characterized in that: chemical formula is C
10+mH
7+nBF
2N
2+xO
y, m wherein, n, x, y are the integer of 0-100, and molecule has following structure:
Wherein: R
1, R
2, R
3Be alkyl, aryl, thiazolinyl, alkynyl, ester group, ether, thioether group, cyanic acid or halogen atom; R
4Be alkyl, aryl, ether, hydroxyl or amino; R
5, R
6Be alkyl, alkoxyl group, ether, thioether group, alkynyl or fluorine atom;
Be negative charge,
Be positive charge.
2. preparation method of novel red light BODIPY optical dye according to claim 1 is characterized in that step is following:
1) will have substituent R
1, R
2, R
3The pyrroles be dissolved in the organic solvent, under nitrogen atmosphere, add oxoethanoic acid second fat, obtain mixed solution, in the mixed solution organic solvent with have substituent R
1, R
2, R
3Pyrroles's volume ratio be 1-10000:1, oxoethanoic acid second fat with have substituent R
1, R
2, R
3Pyrroles's mol ratio be 0.01-0.50:1;
2) under nitrogen protection, be added to catalyst drops in the reaction solution, be 0-100 ℃ in temperature and reacted 3 hours down, obtain reaction solution;
3) in above-mentioned reaction solution, add 2,3-two chloro-5,6-dicyano-1,4-benzoquinones (DDQ), the mol ratio of DDQ and oxoethanoic acid second fat is 0.01-10.0:1, oxydehydrogenation 0.5 hour obtains dehydrogenation liquid;
4) in above-mentioned dehydrogenation liquid, add organic amine and boron trifluoride ether solution successively; 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) after underpressure distillation removed and desolvates, residue obtained target compound through silica gel column chromatography.
3. according to the preparation method of the said novel red light BODIPY optical dye of claim 2, it is characterized in that: the described substituting group that has is R
1, R
2, R
3The pyrroles in substituent R
1, R
2, R
3For alkyl, aryl, thiazolinyl, alkynyl, ester group, ether, thioether group, cyanic acid or halogen atom and be directly connected on the pyrrole ring.
4. according to the preparation method of the said novel red light BODIPY optical dye of claim 2, it is characterized in that: said organic solvent is methylene dichloride, chloroform, benzene,toluene,xylene, ether, THF, ethyl acetate, methyl-sulphoxide or dioxane.
5. according to the preparation method of the said novel red light BODIPY optical dye of claim 2, it is characterized in that: said catalyzer is trifluoroacetic acid or tosic acid.
6. according to the preparation method of the said novel red light BODIPY optical dye of claim 2, it is characterized in that: said organic amine is triethylamine, Trimethylamine 99, diisopropylethylamine, diisopropyl aniline or piperidines.
7. application of novel red light BODIPY optical dye according to claim 1; It is characterized in that: be used for cell imaging, fluorescent probe or laser dyes, method is: can this luminescent dye molecule be dissolved in the aqueous buffer solution of saline water or HEPES (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 directly is used for various application scenarios to the above-mentioned solution that is dissolved with this optical dye then.
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| CN113912632A (en) * | 2021-11-10 | 2022-01-11 | 四川大学华西医院 | Novel fluoroboric fluorescent dye and preparation method and application 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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