CN114805262B - Viscosity and polarity response type platform fluorescent probe, hydrogen sulfide detection fluorescent probe, and synthesis process and application thereof - Google Patents
Viscosity and polarity response type platform fluorescent probe, hydrogen sulfide detection fluorescent probe, and synthesis process and application thereof Download PDFInfo
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- CN114805262B CN114805262B CN202210483797.XA CN202210483797A CN114805262B CN 114805262 B CN114805262 B CN 114805262B CN 202210483797 A CN202210483797 A CN 202210483797A CN 114805262 B CN114805262 B CN 114805262B
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- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 104
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 title claims abstract description 78
- 229910000037 hydrogen sulfide Inorganic materials 0.000 title claims abstract description 78
- 238000001514 detection method Methods 0.000 title claims abstract description 49
- 238000000034 method Methods 0.000 title claims abstract description 24
- 230000004044 response Effects 0.000 title claims abstract description 22
- 230000008569 process Effects 0.000 title claims abstract description 14
- 230000015572 biosynthetic process Effects 0.000 title claims abstract description 8
- 238000003786 synthesis reaction Methods 0.000 title claims abstract description 8
- 230000007613 environmental effect Effects 0.000 claims abstract description 8
- 230000002194 synthesizing effect Effects 0.000 claims abstract description 6
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 39
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 38
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 35
- 238000006243 chemical reaction Methods 0.000 claims description 30
- 239000002904 solvent Substances 0.000 claims description 24
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 15
- LOTKRQAVGJMPNV-UHFFFAOYSA-N 1-fluoro-2,4-dinitrobenzene Chemical compound [O-][N+](=O)C1=CC=C(F)C([N+]([O-])=O)=C1 LOTKRQAVGJMPNV-UHFFFAOYSA-N 0.000 claims description 12
- 125000001424 substituent group Chemical group 0.000 claims description 12
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 9
- XFVZSRRZZNLWBW-UHFFFAOYSA-N 4-(Diethylamino)salicylaldehyde Chemical compound CCN(CC)C1=CC=C(C=O)C(O)=C1 XFVZSRRZZNLWBW-UHFFFAOYSA-N 0.000 claims description 8
- 150000001875 compounds Chemical class 0.000 claims description 8
- 238000010438 heat treatment Methods 0.000 claims description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 6
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 6
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 6
- 239000002994 raw material Substances 0.000 claims description 6
- 238000001727 in vivo Methods 0.000 claims description 5
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 4
- 239000003513 alkali Substances 0.000 claims description 3
- 239000002585 base Substances 0.000 claims description 3
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 3
- 238000006555 catalytic reaction Methods 0.000 claims description 2
- 238000003745 diagnosis Methods 0.000 claims 1
- 201000010099 disease Diseases 0.000 claims 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims 1
- PYLWMHQQBFSUBP-UHFFFAOYSA-N monofluorobenzene Chemical compound FC1=CC=CC=C1 PYLWMHQQBFSUBP-UHFFFAOYSA-N 0.000 claims 1
- 239000000523 sample Substances 0.000 abstract description 49
- 230000008859 change Effects 0.000 abstract description 13
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- 239000000243 solution Substances 0.000 description 34
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- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 9
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- 239000012491 analyte Substances 0.000 description 9
- 239000012295 chemical reaction liquid Substances 0.000 description 9
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- 239000000872 buffer Substances 0.000 description 7
- 229910052979 sodium sulfide Inorganic materials 0.000 description 7
- GRVFOGOEDUUMBP-UHFFFAOYSA-N sodium sulfide (anhydrous) Chemical compound [Na+].[Na+].[S-2] GRVFOGOEDUUMBP-UHFFFAOYSA-N 0.000 description 7
- 239000000203 mixture Substances 0.000 description 6
- 238000002390 rotary evaporation Methods 0.000 description 6
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- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 4
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 4
- 150000001450 anions Chemical class 0.000 description 4
- 238000012512 characterization method Methods 0.000 description 4
- 235000019439 ethyl acetate Nutrition 0.000 description 4
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- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 3
- 235000018417 cysteine Nutrition 0.000 description 3
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 3
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- CMDGQTVYVAKDNA-UHFFFAOYSA-N propane-1,2,3-triol;hydrate Chemical compound O.OCC(O)CO CMDGQTVYVAKDNA-UHFFFAOYSA-N 0.000 description 3
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- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 2
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 2
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 2
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 2
- 235000003704 aspartic acid Nutrition 0.000 description 2
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 2
- 150000001768 cations Chemical class 0.000 description 2
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- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 2
- 125000001183 hydrocarbyl group Chemical group 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
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- 150000003573 thiols Chemical class 0.000 description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 2
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
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- 230000000916 dilatatory effect Effects 0.000 description 1
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- 238000011065 in-situ storage Methods 0.000 description 1
- 239000002440 industrial waste Substances 0.000 description 1
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- 230000003914 insulin secretion Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 229940046892 lead acetate Drugs 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000034217 membrane fusion Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000004770 neurodegeneration Effects 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000010534 nucleophilic substitution reaction Methods 0.000 description 1
- 230000035778 pathophysiological process Effects 0.000 description 1
- 239000002798 polar solvent Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
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- 102000004196 processed proteins & peptides Human genes 0.000 description 1
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- 230000019491 signal transduction Effects 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000002341 toxic gas Substances 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D307/00—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom
- C07D307/77—Heterocyclic compounds containing five-membered rings having one oxygen atom as the only ring hetero atom ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D307/78—Benzo [b] furans; Hydrogenated benzo [b] furans
- C07D307/82—Benzo [b] furans; Hydrogenated benzo [b] furans with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to carbon atoms of the hetero ring
- C07D307/84—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1088—Heterocyclic compounds characterised by ligands containing oxygen as the only heteroatom
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6432—Quenching
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- Organic Chemistry (AREA)
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- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
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- Optics & Photonics (AREA)
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- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a viscosity and polarity response type platform fluorescent probe, a hydrogen sulfide detection fluorescent probe, a synthesis process and application thereof. The structure of the hydrogen sulfide detection fluorescent probe is shown as follows;. Aiming at the problems of poor operability, low sensitivity and the like in the detection of polarity and viscosity in a physiological microenvironment, the invention provides a high-sensitivity platform fluorescent probe for detecting the environmental viscosity and the polarity, which can react to the change of the environmental viscosity through the severe change of the fluorescence intensity and the change of the environmental polarity through the change of the fluorescence emission position; meanwhile, the probe molecule can serve as an excellent platform molecule for synthesizing probes in different application fields; the high-sensitivity hydrogen sulfide detection fluorescent probe is obtained by modifying the hydroxyl of the fluorescent probe molecule, the detection sensitivity can reach 36 nanomoles, and the fluorescent probe can be used for monitoring hydrogen sulfide in cells.
Description
Technical Field
The invention relates to the technical field of in-vivo hydrogen sulfide detection, in particular to a viscosity and polarity response type platform fluorescent probe, a hydrogen sulfide detection fluorescent probe, a synthesis process and application thereof.
Background
The information disclosed in the background of the invention is only for enhancement of understanding of the general background of the invention and is not necessarily to be taken as an admission or any form of suggestion that this information forms the prior art already known to a person of ordinary skill in the art.
Viscosity and polarity are two important indexes in biological microenvironment, and are closely related to a plurality of pathophysiological processes such as inflammation, neurodegenerative diseases, cancers and the like. On the one hand, the viscosity change in a biological system can influence signal transduction, biological macromolecule interaction, apoptosis, autophagy and other processes of cells; on the other hand, many physiological processes such as protein denaturation, polypeptide aggregation, membrane fusion and conformational changes of enzymes are affected by changes in the polarity of the physiological environment. However, the measurement of viscosity and polarity in physiological microenvironments is still an important challenge in chemical biology.
Hydrogen sulfide is a toxic gas in industrial waste gas and is also an important endogenous signal gas transmitter in life systems. In vivo, cysteine and its derivatives can be metabolized by enzymes to produce hydrogen sulfide. Research shows that endogenous hydrogen sulfide participates in the physiological process of cells and has various physiological functions of dilating blood vessels, regulating blood pressure, regulating insulin secretion and the like. The traditional hydrogen sulfide detection method comprises a standard iodine method, a lead acetate test paper method, an ion activity method, a hydrogen sulfide instrument method and the like. However, the conventional analysis method cannot perform in-vivo detection, and has the defects of dependence on a large detection instrument, complex pretreatment of a sample to be detected, low detection speed, inapplicability to real-time in-situ detection and the like.
Disclosure of Invention
Aiming at the problems, the invention provides a viscosity and polarity response type platform fluorescent probe, a hydrogen sulfide detection fluorescent probe, a synthesis process and application thereof, and the hydrogen sulfide detection fluorescent probe constructed based on the fluorescent probe has the functions of high selectivity and high sensitivity for detecting hydrogen sulfide and can be used for monitoring hydrogen sulfide in cells in real time. In order to achieve the above object, the technical scheme of the present invention is as follows:
in a first aspect of the invention, the invention discloses a viscosity and polarity responsive platform fluorescent probe, the structure of which is shown in formula 3.
In the above formula 3, the substituent R 1 Is hydrocarbonRadicals such as methyl, ethyl, and the like; substituent R 2 is-CN, -CONH 2 -COOH, etc.
In a second aspect of the invention, the invention discloses a hydrogen sulfide detection fluorescent probe, the structure of which is shown in formula 4.
In the above formula 4, the substituent R 1 Is a hydrocarbon group such as methyl, ethyl, etc.; substituent R 2 is-CN, -CONH 2 -COOH, etc.
In a third aspect of the invention, the invention discloses a synthesis process of the viscosity and polarity response type platform fluorescent probe, which comprises the following steps: the method comprises the steps of taking a compound shown in a formula 1 and 4-diethylaminosalicylaldehyde as raw materials, heating and reacting, and separating out a target product shown in a formula 3.
In the formula 1, the substituent R 1 Is a hydrocarbon group, e.g. methyl (-CH) 3 ) Ethyl (-CH) 2 CH 3 ) Etc.; substituent R 2 is-CN, -CONH 2 -COOH, etc.
Further, the molar ratio of the compound shown in the formula 1 to the 4-diethylaminosalicylaldehyde is in the range of 1:1 to 4:1.
further, the compound shown in the formula 1 and 4-diethylaminosalicylaldehyde are dissolved in a solvent and then heated for reaction. Optionally, the solvent comprises: methanol, ethanol, N-Dimethylformamide (DMF), dimethyl sulfoxide (DMSO), and the like.
Further, the temperature of the heating reaction is 50-100 ℃.
In a fourth aspect of the present invention, the present invention discloses a synthesis process of the hydrogen sulfide detection fluorescent probe, comprising the steps of: the preparation method comprises the steps of taking a viscosity and polarity response type platform fluorescent probe shown in a formula 3 and 2, 4-dinitrofluorobenzene as raw materials, and separating a target product formula 4 after reaction under the catalysis of alkali.
Further, the molar ratio of the viscosity and polarity responsive platform fluorescent probe of formula 3 to 2, 4-dinitrofluorobenzene is in the range of 1:1 to 1:1.5.
further, the viscosity and polarity responsive platform fluorescent probe of formula 3 is reacted with 2, 4-dinitrofluorobenzene after being dissolved in a solvent. Optionally, the solvent comprises: dichloromethane, chloroform, acetonitrile, and the like.
Further, the addition ratio of the base is one time or more of the molar amount of the raw material represented by formula 3. Preferably, the base includes any one of triethylamine, potassium carbonate, and the like.
In a fifth aspect of the invention, the viscosity and polarity responsive platform fluorescent probe (formula 3) and the hydrogen sulfide detection fluorescent probe (formula 4) are disclosed to be applied to the fields of biology, medicine and the like, and are preferably used for monitoring in vivo hydrogen sulfide.
Compared with the prior art, the invention has at least the following beneficial effects:
aiming at the problems of poor operability, low sensitivity and the like in the detection of polarity and viscosity in a physiological microenvironment, the invention provides a high-sensitivity platform fluorescent probe (formula 3) which can be used for detecting the environmental viscosity and the polarity, and the probe can react to the change of the environmental viscosity through the severe change of the fluorescence intensity and the change of the environmental polarity through the change of the fluorescence emission position; meanwhile, the probe molecule can serve as an excellent platform molecule for synthesizing probes in different application fields; the hydroxyl of the fluorescent probe molecule is modified to obtain a high-sensitivity hydrogen sulfide detection fluorescent probe (formula 4), the fluorescence of the molecule is quenched by adopting a 2, 4-dinitrofluorobenzene unit containing a strong electron-withdrawing nitro group, and the hydrogen sulfide molecule can undergo aromatic nucleophilic substitution reaction with the unit to cause the 2, 4-dinitrofluorobenzene unit to be separated from the hydrogen sulfide fluorescent probe (formula 4) molecule, so that a molecular fluorescent signal is recovered, and the detection of hydrogen sulfide is realized. Through tests, the detection sensitivity of the fluorescent probe for detecting hydrogen sulfide is as high as 36 nanomoles, and the fluorescent probe can be used for monitoring hydrogen sulfide in a special environment of cells.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention.
FIG. 1 is a nuclear magnetic resonance hydrogen spectrum of a viscosity and polarity responsive platform fluorescent probe molecule synthesized according to a first embodiment of the present invention.
FIG. 2 is an enlarged view showing the nuclear magnetic resonance hydrogen spectrum of the viscosity and polarity responsive platform fluorescent probe molecule synthesized in the first embodiment of the present invention in the region of chemical shift 6.0-8.8.
FIG. 3 is a nuclear magnetic resonance hydrogen spectrum of a hydrogen sulfide detecting fluorescent probe synthesized according to a third embodiment of the present invention.
FIG. 4 is a graph showing fluorescence emission of the viscosity and polarity responsive platform fluorescent probe molecules synthesized according to the first embodiment of the present invention in different viscosity systems composed of glycerol-water.
FIG. 5 is a graph showing the quantitative relationship between the fluorescence intensity and viscosity of the viscosity and polarity responsive platform fluorescent probe molecules synthesized in the first example of the present invention in different viscosity systems composed of glycerol-water.
FIG. 6 is a graph showing fluorescence emission of viscosity and polarity responsive platform fluorescent probe molecules synthesized in accordance with the first embodiment of the present invention in solvents of different polarities.
FIG. 7 is a graph showing the quantitative relationship between the viscosity and the fluorescence emission wavelength of polar-responsive platform fluorescent probe molecules in solvents of different polarities, synthesized in accordance with the first embodiment of the present invention.
FIG. 8 is a fluorescence spectrum of a hydrogen sulfide detecting fluorescent probe synthesized according to the second embodiment of the present invention after respective reactions with 28 kinds of analytes.
FIG. 9 is a graph showing fluorescence intensity at 530nm after a hydrogen sulfide detection fluorescent probe synthesized according to the second embodiment of the present invention reacts with 28 kinds of analytes, respectively.
FIG. 10 is a graph showing the fluorescence intensity of a hydrogen sulfide detecting fluorescent probe synthesized according to the second embodiment of the present invention at 530nm after reacting with sodium sulfide in the presence of various interferents.
FIG. 11 is a graph showing the fluorescence intensity of a hydrogen sulfide detecting fluorescent probe synthesized according to the second embodiment of the present invention at various time points when it reacts with sodium sulfide.
FIG. 12 is a graph showing fluorescence intensity of a hydrogen sulfide detection fluorescent probe synthesized according to the second embodiment of the present invention after reacting with hydrogen sulfide of different concentrations for the same period of time.
FIG. 13 is a graph showing the relationship between fluorescence intensity and hydrogen sulfide concentration after the same time period as the reaction of hydrogen sulfide with a hydrogen sulfide detection fluorescent probe synthesized in the second embodiment of the present invention.
FIG. 14 is a graph showing the results of cell viability after treatment of HeLa cells with different concentrations of the second example synthesized hydrogen sulfide detecting fluorescent probe.
FIG. 15 is a graph showing the results of fluorescence confocal imaging applications of hydrogen sulfide detection fluorescent probes synthesized according to the second embodiment of the present invention to hydrogen sulfide in cells.
Detailed Description
In the following description, specific details of the invention are set forth in order to provide a thorough understanding of the invention. The terminology used in the description of the invention herein is for the purpose of describing the advantages and features of the invention only and is not intended to be limiting of the invention.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The medicines or reagents used in the present invention are used according to the product instructions or by the conventional methods of use in the art unless specifically stated. The process according to the invention will now be further described with reference to the drawings and the detailed description.
First embodiment
A method for preparing a viscosity and polarity responsive platform fluorescent probe, referring to scheme 1, comprising the steps of:
(1) 4mmol of 4-diethylaminosalicylaldehyde (formula 2), 1mmol of the compound represented by formula 1, and 20mL of methanol were weighed into a three-necked flask, and the above-mentioned system was heated at 50℃to react.
(2) TLC monitoring reaction, cooling the reaction liquid to room temperature when the reaction intermediate disappears, removing part of solvent by rotary evaporation, and separating out a large amount of crystals after the reaction liquid is cooled; and (3) filtering the solid-liquid mixture to obtain an orange solid, namely the viscosity and polarity response type platform fluorescent probe, as shown in a formula 3.
The nuclear magnetic resonance hydrogen spectra of the viscosity and polarity responsive platform fluorescent probe of formula 3 synthesized in this example are shown in fig. 1 and 2. Wherein FIG. 1 is a nuclear magnetic resonance hydrogen spectrum of the fluorescent probe molecule, and FIG. 2 is an enlarged view of the nuclear magnetic resonance hydrogen spectrum of the fluorescent probe molecule in a region with a chemical shift of 6.0-8.8. From FIGS. 1 and 2, it can be determined that the viscosity and polarity responsive platform fluorescent probe is shown in formula 3.
The structural characterization data of the viscosity and polarity responsive platform fluorescent probe synthesized in this example are as follows: 1 HNMR(500MHz,DMSO-d 6 ),δ(ppm):12.39(s,1H),8.75(d,J＝1.55Hz,1H),8.01(dd,J 1 ＝8.60Hz,J 2 ＝1.70Hz,1H),7.41(d,J＝8.55Hz,1H),7.20(d,J＝9.00Hz,1H),6.33(dd,J 1 ＝8.95Hz,J 2 ＝1.90Hz 1H),6.13(d,J＝2.00Hz,1H),4.40(q,J＝7.15Hz,2H),3.42(q,J＝7.15Hz,4H),1.43(t,J＝7.15Hz,3H),1.23(t,J＝7.15Hz,6H)；ESI-MS:m/z[M+H] + calcd for:C 23 H 24 N 3 O 4 :406.1；found 406.1.ESI-MS:m/z[M+H] + calcd for:C 29 H 26 N 5 O 8 :572.1；found 572.2.
second embodiment
A preparation method of a viscosity and polarity response type platform fluorescent probe, referring to the reaction route 1, comprises the following steps:
(1) 1mmol of 4-diethylaminosalicylaldehyde (formula 2), 1mmol of the compound of formula 1 and 5mL of DMF were weighed into a three-necked flask, and the above-mentioned system was heated at 100 ℃.
(2) TLC monitoring reaction, cooling the reaction liquid to room temperature when the reaction intermediate disappears; slowly adding the reaction solution into 50mL of saturated saline water, and precipitating a large amount of solids; and (3) filtering the solid-liquid mixture to obtain an orange solid, namely the viscosity and polarity response type platform fluorescent probe, as shown in a formula 3.
The structural characterization data of the viscosity and polarity responsive platform fluorescent probe synthesized in this example are as follows: 1 HNMR(500MHz,DMSO-d 6 ),δ(ppm):12.39(s,1H),8.75(d,J＝1.55Hz,1H),8.01(dd,J 1 ＝8.60Hz,J 2 ＝1.70Hz,1H),7.41(d,J＝8.55Hz,1H),7.20(d,J＝9.00Hz,1H),6.33(dd,J 1 ＝8.95Hz,J 2 ＝1.90Hz,1H),6.13(d,J＝2.00Hz,1H),4.40(q,J＝7.15Hz,2H),3.42(q,J＝7.15Hz,4H),1.43(t,J＝7.15Hz,3H),1.23(t,J＝7.15Hz,6H)；ESI-MS:m/z[M+H] + calcd for:C 23 H 24 N 3 O 4 :406.1；found 406.1.
third embodiment
A method for preparing a hydrogen sulfide detection fluorescent probe, referring to reaction scheme 2, comprising the steps of:
(1) Weighing 0.5mmol of the viscosity and polarity response type platform fluorescent probe of formula 3 synthesized in the first embodiment, 0.6mmol of 2, 4-dinitrofluorobenzene, 0.55mmol of triethylamine and 10mL of dichloromethane in a three-necked flask; the mixture was heated by reaction at 35 ℃.
(2) TLC monitors that the reaction is complete, the reaction liquid is cooled to room temperature, the reaction liquid is extracted and washed three times by deionized water, the organic layer is dried by anhydrous sodium sulfate, and the solvent is removed by rotary evaporation to obtain a crude product. Recrystallizing the crude product by using a mixed solution of ethanol and dichloromethane (ethanol: dichloromethane=1:1), and carrying out suction filtration to obtain a red solid, namely the hydrogen sulfide detection fluorescent probe, as shown in the formula 4.
The nuclear magnetic resonance hydrogen spectrum of the hydrogen sulfide detection fluorescent probe represented by formula 4 synthesized in this example is shown in FIG. 3, and the structural formula 4 of the hydrogen sulfide detection fluorescent probe can be determined from FIG. 3.
The embodiment is combined withThe structural characterization data of the viscosity and polarity response type platform fluorescent probe are as follows: 1 HNMR(500MHz,DMSO-d 6 ),δ(ppm):8.94(s,1H),8.92(d,J＝2.75Hz,1H),8.35(dd,J 1 ＝9.20Hz,J 2 ＝2.75Hz,1H),8.28(d,J＝1.75Hz,1H),8.25(d,J＝9.15Hz,1H),8.03(dd,J 1 ＝8.70Hz,J 2 ＝1.80Hz,1H),7.43(d,J＝8.60Hz,1H),7.10(d,J＝9.30Hz,1H),6.73(dd,J 1 ＝9.15Hz,J 2 ＝2.50Hz,1H),6.25(d,J＝2.50Hz,1H),4.43(q,J＝7.10Hz,2H),3.49(q,J＝7.05Hz,4H),1.44(t,J＝7.15Hz,3H),1.27(t,J＝6.95Hz,6H)；ESI-MS:m/z[M+H] + calcd for:C 29 H 26 N 5 O 8 :572.1；found 572.2.
fourth embodiment
A preparation method of a hydrogen sulfide detection fluorescent probe, referring to the reaction route 2, comprises the following steps:
(1) Weighing 0.5mmol of the viscosity and polarity response type platform fluorescent probe shown in formula 3 synthesized in the first embodiment, 0.6mmol of 2, 4-dinitrofluorobenzene, 0.55mmol of triethylamine and 10mL of acetonitrile in a three-necked flask; the mixture was heated by reaction at 35 ℃.
(2) TLC monitored completion of the reaction, stopped the reaction, removed the solvent by rotary evaporation, washed three times with 10mL of dichloromethane solution, saturated brine, the organic layer dried over anhydrous sodium sulfate, and removed the solvent by rotary evaporation to give the crude product. Recrystallizing the crude product by using a mixed solution of ethanol and dichloromethane (ethanol: dichloromethane=1:1), and carrying out suction filtration to obtain a red solid, namely the hydrogen sulfide detection fluorescent probe, as shown in the formula 4.
The structural characterization data of the viscosity and polarity responsive platform fluorescent probe synthesized in this example are as follows: 1 HNMR(500MHz,DMSO-d 6 ),δ(ppm):8.94(s,1H),8.92(d,J＝2.75Hz,1H),8.35(dd,J 1 ＝9.20Hz,J 2 ＝2.75Hz,1H),8.28(d,J＝1.75Hz,1H),8.25(d,J＝9.15Hz,1H),8.03(dd,J 1 ＝8.70Hz,J 2 ＝1.80Hz,1H),7.43(d,J＝8.60Hz,1H),7.10(d,J＝9.30Hz,1H),6.73(dd,J 1 ＝9.15Hz,J 2 ＝2.50Hz,1H),6.25(d,J＝2.50Hz,1H),4.43(q,J＝7.10Hz,2H),3.49(q,J＝7.05Hz,4H),1.44(t,J＝7.15Hz,3H),1.27(t,J＝6.95Hz,6H)；ESI-MS:m/z[M+H] + calcd for:C 29 H 26 N 5 O 8 :572.1；found 572.2.
fifth embodiment
A method for preparing a hydrogen sulfide detection fluorescent probe, referring to reaction scheme 3, comprising the steps of:
(1) Weighing 0.5mmol of the viscosity and polarity responsive platform fluorescent probe of formula 3 synthesized in the first example, 0.5mmol of 2, 4-dinitrofluorobenzene, 0.5mmol of potassium carbonate and 10mL of dichloromethane in a three-necked flask; the mixture was heated by reaction at 35 ℃.
(2) TLC monitors that the reaction is complete, the reaction liquid is cooled to room temperature, the reaction liquid is extracted and washed three times by deionized water, the organic layer is dried by anhydrous sodium sulfate, and the solvent is removed by rotary evaporation to obtain a crude product. Recrystallizing the crude product by using a mixed solution of ethanol and dichloromethane (ethanol: dichloromethane=1:1), and performing suction filtration to obtain a red solid, namely the hydrogen sulfide detection fluorescent probe, wherein the nuclear magnetic resonance hydrogen spectrum display structure of the red solid is shown as a formula 5.
Sixth embodiment
A method for preparing a hydrogen sulfide detection fluorescent probe, referring to reaction scheme 4, comprising the steps of:
(1) Weighing 0.6mmol of the viscosity and polarity response type platform fluorescent probe of formula 3 synthesized in the first example, 0.9mmol of 2, 4-dinitrofluorobenzene, 0.9mmol of triethylamine and 10mL of chloroform in a three-necked flask; the mixture was heated by reaction at 35 ℃.
(2) TLC monitors that the reaction is complete, the reaction liquid is cooled to room temperature, the reaction liquid is extracted and washed three times by deionized water, the organic layer is dried by anhydrous sodium sulfate, and the solvent is removed by rotary evaporation to obtain a crude product. Recrystallizing the crude product by using a mixed solution of ethanol and dichloromethane (ethanol: dichloromethane=1:1), and performing suction filtration to obtain a red solid, namely the hydrogen sulfide detection fluorescent probe, wherein the nuclear magnetic resonance hydrogen spectrum display structure of the red solid is shown as a formula 6.
Performance index test
1. Weighing the viscosity and polarity response type platform fluorescent probe molecules (formula 3) synthesized in the first embodiment, dissolving the probe molecules in DMSO, and preparing a 1mM probe molecule solution; the above 1mM probe molecule solution was prepared into a 10. Mu.M/L probe solution in a system of different viscosities composed of glycerol-water.
(1) The change in fluorescence intensity in the probe solution was tested and the results are shown in fig. 4. From this fig. 4, it can be seen that the viscosity and polarity responsive platform fluorescent probe molecules exhibit unique sensitivity to different viscosities of the solution system, and the fluorescence intensity increases with increasing system viscosity.
(2) The fluorescence intensity of the probe solution was tested and the system viscosity was linearly fitted, and the results are shown in fig. 5. As can be seen from FIG. 5, the viscosity and the polarity response type platform fluorescent probe molecule have good linear relationship between the fluorescence intensity at 530nm and the viscosity of the system (R 2 =0.99), can be used as an excellent viscosity-changing fluorescent probe.
2. Weighing the viscosity and polarity response type platform fluorescent probe molecules (formula 3) synthesized in the first embodiment, and dissolving the viscosity and polarity response type platform fluorescent probe molecules in DMSO to prepare a 1mM probe molecule solution; the 1mM probe molecule solution is dissolved in solvents of different polarities (including ethyl acetate (EtOAc), methanol (MeOH), ethanol (EtOH), isopropanol (IPA), dimethyl sulfoxide (DMSO), water (H) 2 O)) to prepare a probe solution of 10. Mu.M/L of a solvent of different polarity. The probe solutions of the different polarity solvents were separately tested for fluorescence emission and the results are shown in FIG. 6. From this figure, it can be seen that the probe molecules exhibit unique sensitivity to solvents of different polarity, with different fluorescence emission wavelengths in different solvents.
3. Weigh the viscosity sum synthesized in the first examplePolar response type platform fluorescent probe molecules (formula 3) are dissolved in DMSO to prepare 1mM probe molecule solution; the 1mM probe molecule solution is dissolved in solvents of different polarities (including ethyl acetate (EtOAc), methanol (MeOH), ethanol (EtOH), isopropanol (IPA), dimethyl sulfoxide (DMSO), water (H) 2 O)) was prepared into a probe solution of 10. Mu.M/L of solvents of different polarities.
(1) The quantitative relationship chart of fluorescence emission wavelengths of the probe solutions of the different polar solvents was tested separately, and the results are shown in fig. 7. As can be seen from FIG. 7, the maximum emission wavelength of the viscosity and polarity responsive platform fluorescent probe molecules in these solvents is quantitatively related to the dielectric constant (R 2 =0.99), indicating that the probe molecules can be used for quantitative analysis of environmental polarity changes.
4. The hydrogen sulfide detection fluorescent probe synthesized in the third example (formula 4) was weighed, dissolved in DMSO, and prepared as a 1mM probe molecule solution. The probe molecule solution is taken in Tirs-HCl buffer solution (DMSO/H 2 O=1/1, v/v, ph=7.4), probe buffer formulated at 10 μm/L.
To three identical portions of the probe buffer, analyte was added: cations and anions (Al) 3+ 、Ca 2+ 、Cd 2+ 、Co 2+ 、Cr 3+ 、Cu 2+ 、K + 、Ni 2+ 、Tb 3+ 、Zn 2+ 、Mg 2+ 、S 2 O 3 2- 、SO 3 2- 、SO 4 2- 、HS - 、HSO 3 - 、HSO 4 - 、NO 3 - 、S 2 - Sodium salt of the anion), amino acids (glycine, glutamic acid, arginine, lysine, tyrosine, aspartic acid, histidine), biological thiols (cysteine, glutamic acid), three analyte solutions were obtained, each at a concentration of 200 μm/L.
The three analyte solutions were tested for fluorescence spectra and fluorescence intensities, and the results are shown in fig. 8 and 9. As is clear from fig. 8 and 9, the fluorescence intensity of the analyte solution was significantly changed in the hydrogen sulfide atmosphere, and the fluorescence emission peak was around 530nm, whereas no significant fluorescence change was observed in the other analyte atmospheres. The above experiments confirm that the hydrogen sulfide detection fluorescent probe (formula 4) can be used for specific recognition of hydrogen sulfide molecules.
5. The hydrogen sulfide detection fluorescent probe synthesized in the third example (formula 4) was weighed, dissolved in DMSO, and prepared as a 1mM probe molecule solution. The probe molecule solution is taken in Tirs-HCl buffer solution (DMSO/H 2 O=1/1, v/v, ph=7.4), probe buffer formulated at 10 μm/L.
To three identical portions of the probe buffer, analyte was added: cations and anions (Al) 3+ 、Ca 2+ 、Cd 2+ 、Co 2+ 、Cr 3+ 、Cu 2+ 、K + 、Ni 2+ 、Tb 3+ 、Zn 2+ 、Mg 2+ 、S 2 O 3 2- 、SO 3 2- 、SO 4 2- 、HS - 、HSO 3 - 、HSO 4 - 、NO 3 - 、S 2 - Sodium salt of the anion), amino acids (glycine, glutamic acid, arginine, lysine, tyrosine, aspartic acid, histidine), biological thiols (cysteine, glutamic acid), three analyte solutions were obtained, each at a concentration of 200 μm/L.
200. Mu.M/L sodium sulfide was then added to each of the three analyte solutions. The resulting three analyte solutions (containing sodium sulfide) were tested for fluorescence intensity at 530nm before and after reaction, and the results are shown in FIG. 10. From the figure, the fluorescence probe for detecting hydrogen sulfide (formula 4) does not find significant fluorescence change in the atmosphere of other interferents, but the fluorescence intensity is obviously changed after hydrogen sulfide is added into the interferents, so that the probe molecule can be used for specifically identifying hydrogen sulfide molecules.
6. The hydrogen sulfide detection fluorescent probe synthesized in the third example (formula 4) was weighed, dissolved in DMSO, and prepared as a 1mM probe molecule solution. The probe molecule solution is taken in Tirs-HCl buffer solution (DMSO/H 2 O=1/1, v/v, ph=7.4), probe buffer formulated at 10 μm/L.
200. Mu.M/L sodium sulfide was added to the probe buffer and the fluorescence intensity at various time points of the reaction was tested, and the results are shown in FIG. 11. From this figure, the emission at 530nm almost reaches saturation within 10min, fluorescence is enhanced 95-fold, indicating that the probe molecule responds rapidly to hydrogen sulfide.
7. The hydrogen sulfide detection fluorescent probe synthesized in the third example (formula 4) was weighed, dissolved in DMSO, and prepared as a 1mM probe molecule solution. The probe molecule solution is taken in Tirs-HCl buffer solution (DMSO/H 2 O=1/1, v/v, ph=7.4), probe buffer formulated at 10 μm/L.
Sodium sulfide was added to the probe buffer solution at various concentrations and the fluorescence intensity was measured, and the change in fluorescence intensity of the probe solution with the concentration of hydrogen sulfide was recorded, and the results are shown in fig. 12. From this graph, it is understood that the fluorescence intensity of the probe solution gradually increases with an increase in the concentration of hydrogen sulfide in the range of 2 to 200 μm. The fluorescence intensity of the probe solution was linearly fitted to the concentration of hydrogen sulfide, and the results are shown in FIG. 13. From the graph, the linear relationship exists between the fluorescence intensity of the probe solution and the concentration of hydrogen sulfide in the concentration range of 0-20 μm, and the graph can be used for quantitative detection of hydrogen sulfide.
8. The hydrogen sulfide detection fluorescent probe synthesized in the third example (formula 4) was weighed, dissolved in DMSO, and prepared into a probe molecule mother solution.
Culturing Hela cells with DMEM medium; the survival rate of Hela cells after treatment of the probe molecular mother solution at different concentrations was tested by using the MTT method with the probe molecular mother solution as a mother solution, and the results are shown in FIG. 14. From this figure, it is clear that the hydrogen sulfide detection fluorescent probe (formula 4) is almost nontoxic to cells at a concentration of 0 to 100. Mu.M, and has a high cell survival rate.
9. Hela cells were cultured in DMEM medium for 12 hours using the hydrogen sulfide detection fluorescent probe synthesized in the third example (formula 4), and then incubated in DMEM medium containing 200 μm sodium sulfide for 12 hours, and fluorescence was imaged on a fluorescence confocal microscope, and the results are shown in fig. 15. From the graph, the fluorescent probe has higher signal-to-noise ratio for intracellular hydrogen sulfide, and can be used as a high-contrast imaging probe for detecting exogenous hydrogen sulfide of living cells.
The foregoing is illustrative of only a few embodiments of the present invention and is not to be construed as limiting the scope of the invention. It should be noted that modifications, substitutions, improvements, etc. can be made by others skilled in the art without departing from the spirit and scope of the present invention. The scope of the invention should, therefore, be determined with reference to the appended claims.
Claims (9)
1. A viscosity and polarity response type platform fluorescent probe has a structure shown in a formula 3;
formula 3;
in the formula 3, the substituent R 1 Is ethyl; substituent R 2 is-CN.
2. The process for synthesizing a viscosity and polarity responsive platform fluorescent probe according to claim 1, comprising the steps of: taking a compound shown in a formula 1 and 4-diethylaminosalicylaldehyde as raw materials, and separating a target product shown in a formula 3 after heating reaction;
formula 1;
in the formula 1, the substituent R 1 Is ethyl; substituent R 2 is-CN;
dissolving a compound shown in a formula 1 and 4-diethylaminosalicylaldehyde in a solvent, and then heating for reaction;
the solvent comprises: any one of methanol, ethanol, N-dimethylformamide and dimethyl sulfoxide;
the temperature of the heating reaction is 50-100 ℃.
3. The synthesis process of the viscosity and polarity responsive platform fluorescent probe according to claim 2, wherein the molar ratio of the compound represented by formula 1 to 4-diethylaminosalicylaldehyde is in the range of 1: 1-1: 4.
4. a hydrogen sulfide detection fluorescent probe has a structure shown in a formula 4;
formula 4;
in the formula 4, the substituent R 1 Is ethyl; substituent R 2 is-CN.
5. The process for synthesizing a hydrogen sulfide detecting fluorescent probe as claimed in claim 4, comprising the steps of: taking the viscosity and polarity response type platform fluorescent probe shown in the formula 3 in claim 1 and 2, 4-dinitrofluorobenzene as raw materials, and separating a target product formula 4 after heating reaction under the catalysis of alkali;
the viscosity and polarity response type platform fluorescent probe shown in the formula 3 and 2, 4-dinitrofluorobenzene are dissolved in a solvent and then heated for reaction;
the solvent comprises: any one of dichloromethane, chloroform and acetonitrile;
the temperature of the heating reaction is 35 ℃;
the base comprises any one of triethylamine and potassium carbonate.
6. The process for synthesizing a fluorescent probe for hydrogen sulfide detection according to claim 5, wherein the molar ratio of the viscosity and polarity responsive platform fluorescent probe of formula 3 to 2, 4-dinitrofluorobenzene is in the range of 1: 1-1: 1.5.
7. the process for synthesizing a hydrogen sulfide detecting fluorescent probe according to claim 5, wherein the alkali is added in a proportion of one time or more of the molar amount of the raw material represented by formula 3.
8. The use of a viscosity and polarity responsive platform fluorescent probe according to claim 1, wherein the viscosity and polarity responsive platform fluorescent probe is used for detection of environmental viscosity and polarity.
9. The use of the fluorescent probe for detecting hydrogen sulfide according to claim 4 in the biological and medical fields for the purpose of diagnosis and treatment of non-disease, wherein the fluorescent probe for detecting hydrogen sulfide is used for detecting hydrogen sulfide in vivo.
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| CN111349070A (en) * | 2020-02-12 | 2020-06-30 | 曲阜师范大学 | Near-infrared fluorescent molecular probe for detecting biological cell viscosity and preparation method and application thereof |
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| CN112409430A (en) * | 2019-08-21 | 2021-02-26 | 湖南科技大学 | Fluorescent probe capable of detecting viscosity and hydrogen sulfide, preparation and application thereof |
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