CN114805262A - Viscosity and polarity response type platform fluorescent probe, hydrogen sulfide detection fluorescent probe, synthesis process and application thereof - Google Patents
Viscosity and polarity response type platform fluorescent probe, hydrogen sulfide detection fluorescent probe, synthesis process and application thereof Download PDFInfo
- Publication number
- CN114805262A CN114805262A CN202210483797.XA CN202210483797A CN114805262A CN 114805262 A CN114805262 A CN 114805262A CN 202210483797 A CN202210483797 A CN 202210483797A CN 114805262 A CN114805262 A CN 114805262A
- Authority
- CN
- China
- Prior art keywords
- fluorescent probe
- hydrogen sulfide
- formula
- viscosity
- polarity
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 107
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 title claims abstract description 76
- 229910000037 hydrogen sulfide Inorganic materials 0.000 title claims abstract description 76
- 238000001514 detection method Methods 0.000 title claims abstract description 49
- 230000004044 response Effects 0.000 title claims abstract description 37
- 238000000034 method Methods 0.000 title claims abstract description 23
- 230000008569 process Effects 0.000 title claims abstract description 16
- 230000015572 biosynthetic process Effects 0.000 title claims abstract description 8
- 238000003786 synthesis reaction Methods 0.000 title claims abstract description 8
- 238000012544 monitoring process Methods 0.000 claims abstract description 8
- 230000002194 synthesizing effect Effects 0.000 claims abstract description 8
- 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 32
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
- 239000002904 solvent Substances 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
- 150000001875 compounds Chemical class 0.000 claims description 10
- 125000001424 substituent group Chemical group 0.000 claims description 10
- 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
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 6
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 6
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 6
- 125000002496 methyl group Chemical group [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
- 125000001183 hydrocarbyl group Chemical group 0.000 claims description 5
- 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
- 238000010438 heat treatment Methods 0.000 claims description 4
- 239000003513 alkali Substances 0.000 claims description 3
- 239000002585 base Substances 0.000 claims description 3
- 238000006555 catalytic reaction Methods 0.000 claims description 3
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 3
- 239000000523 sample Substances 0.000 abstract description 46
- 230000008859 change Effects 0.000 abstract description 14
- 230000007613 environmental effect Effects 0.000 abstract description 7
- 230000035945 sensitivity Effects 0.000 abstract description 6
- 125000002887 hydroxy group Chemical group [H]O* 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 38
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 10
- 239000001257 hydrogen Substances 0.000 description 10
- 229910052739 hydrogen Inorganic materials 0.000 description 10
- 239000007787 solid Substances 0.000 description 9
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 8
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 8
- 239000012043 crude product Substances 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- 238000001228 spectrum Methods 0.000 description 8
- 239000012491 analyte Substances 0.000 description 7
- 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
- 238000005303 weighing Methods 0.000 description 7
- 238000002390 rotary evaporation Methods 0.000 description 6
- 238000000967 suction filtration Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 238000005481 NMR spectroscopy Methods 0.000 description 5
- 239000007853 buffer solution Substances 0.000 description 5
- 239000012295 chemical reaction liquid Substances 0.000 description 5
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- 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
- 238000001816 cooling Methods 0.000 description 4
- 235000019439 ethyl acetate Nutrition 0.000 description 4
- 235000013922 glutamic acid Nutrition 0.000 description 4
- 239000004220 glutamic acid Substances 0.000 description 4
- 239000011259 mixed solution Substances 0.000 description 4
- 239000012044 organic layer Substances 0.000 description 4
- 239000002798 polar solvent Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 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
- 239000008367 deionised water Substances 0.000 description 3
- 229910021641 deionized water Inorganic materials 0.000 description 3
- 239000012452 mother liquor Substances 0.000 description 3
- 230000035790 physiological processes and functions Effects 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000004475 Arginine Substances 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 2
- 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
- 239000007806 chemical reaction intermediate Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000002189 fluorescence spectrum Methods 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 2
- 150000002431 hydrogen Chemical class 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 2
- CMDGQTVYVAKDNA-UHFFFAOYSA-N propane-1,2,3-triol;hydrate Chemical compound O.OCC(O)CO CMDGQTVYVAKDNA-UHFFFAOYSA-N 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 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
- 238000005406 washing Methods 0.000 description 2
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 239000004215 Carbon black (E152) Substances 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
- 238000004220 aggregation Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000004900 autophagic degradation Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000010226 confocal imaging Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 239000002440 industrial waste Substances 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000003914 insulin secretion Effects 0.000 description 1
- 230000003993 interaction 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
- 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
- 239000010413 mother solution Substances 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
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000002040 relaxant effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 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
Images
Classifications
-
- 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
-
- 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"
-
- 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
-
- 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
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- 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)
- Materials Engineering (AREA)
- Optics & Photonics (AREA)
- Engineering & Computer Science (AREA)
- 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 polarity and viscosity detection in a physiological microenvironment, the invention provides a high-sensitivity platform fluorescent probe for detecting environmental viscosity and polarity, which can react the change of environmental viscosity through the drastic change of fluorescence intensity and the change of environmental polarity through the change of a fluorescence emission position; at the same time, the probe molecule can be used as a good probeThe platform molecule is used for synthesizing probes in different application fields; the hydroxyl of the fluorescent probe molecule is modified to obtain the high-sensitivity hydrogen sulfide detection fluorescent probe, the detection sensitivity of the high-sensitivity hydrogen sulfide detection fluorescent probe can reach 36 nanomole, and the high-sensitivity hydrogen sulfide detection 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 this 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 acknowledgement or any form of suggestion that this information forms the prior art already known to a person skilled 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 and even cancer. On one hand, the change of viscosity in a biological system can affect the processes of signal transduction, biomacromolecule interaction, apoptosis, autophagy and the like 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, measurement of viscosity and polarity in physiological microenvironments remains a significant challenge in chemical biology today.
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 enzyme catalysis to produce hydrogen sulfide. Research shows that endogenous hydrogen sulfide is involved in the physiological process of cells and has various physiological functions of relaxing blood vessels, regulating blood pressure, regulating insulin secretion and the like. The traditional detection method of hydrogen sulfide comprises a standard iodine amount 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 disadvantages of dependence on a large-scale detection instrument, complex pretreatment of a sample to be detected, slow 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. In order to achieve the purpose, the technical scheme of the invention is as follows:
in a first aspect of the invention, the invention discloses a viscosity and polarity response type platform fluorescent probe, which has a structure shown in formula 3.
In the above formula 3, the substituent R 1 Is a hydrocarbon group such as methyl, ethyl, etc.; substituent R 2 is-CN, -CONH 2 and-COOH, etc.
In a second aspect of the invention, the invention discloses a hydrogen sulfide detection fluorescent probe, which has a structure 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 and-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: taking a compound shown in a formula 1 and 4-diethylamino salicylaldehyde as raw materials, heating to react, and separating out a target product shown in a formula 3 to obtain the compound.
In the formula 1, the substituent R 1 Is a hydrocarbon radical, e.g. methyl (-CH) 3 ) Ethyl (-CH) 2 CH 3 ) Etc.; substituent R 2 is-CN, -CONH 2 and-COOH, etc.
Further, the molar ratio of the compound shown in the formula 1 to the 4-diethylamino salicylaldehyde is in a range of 1: 1-4: 1.
further, the compound represented by formula 1 and 4-diethylamino salicylaldehyde are dissolved in a solvent and then heated for reaction. Optionally, the solvent comprises: methanol, ethanol, N-Dimethylformamide (DMF), dimethyl sulfoxide (DMSO), etc.
Further, the temperature of the heating reaction is 50-100 ℃.
In a fourth aspect of the invention, the invention discloses a synthesis process of the hydrogen sulfide detection fluorescent probe, which comprises the following steps: taking a viscosity and polarity response type platform fluorescent probe shown in a formula 3 and 2, 4-dinitrofluorobenzene as raw materials, reacting under the catalysis of alkali, and separating out a target product shown in a formula 4 to obtain the fluorescent probe.
Further, the molar ratio of the viscosity and polarity response type platform fluorescent probe shown in formula 3 to 2, 4-dinitrofluorobenzene is in the range of 1: 1-1: 1.5.
further, the viscosity and polarity response type platform fluorescent probe shown in formula 3 and 2, 4-dinitrofluorobenzene are dissolved in a solvent and then react. Optionally, the solvent comprises: dichloromethane, chloroform, acetonitrile, and the like.
Further, the alkali is added in a proportion of one time or more than 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 the fifth aspect of the invention, the application of the viscosity and polarity response type platform fluorescent probe (formula 3) and the hydrogen sulfide detection fluorescent probe (formula 4) in the fields of biology, medicine and the like is disclosed, and the application is preferably used for monitoring hydrogen sulfide in vivo.
Compared with the prior art, the invention has the following beneficial effects:
aiming at the problems of poor operability, low sensitivity and the like in polarity and viscosity detection in a physiological microenvironment, the invention provides a high-sensitivity platform fluorescent probe (formula 3) for detecting environmental viscosity and polarity, which can react the change of environmental viscosity through the drastic change of fluorescence intensity and the change of environmental polarity through the change of a fluorescence emission position; meanwhile, the probe molecule can be used as an excellent platform molecule for synthesizing probes in different application fields; the hydroxyl of the fluorescent probe molecule is reformed to obtain the high-sensitivity hydrogen sulfide detection fluorescent probe (formula 4), the 2, 4-dinitrofluorobenzene unit containing strong electron-withdrawing nitro is adopted to quench molecular fluorescence, and the hydrogen sulfide molecule and the unit can generate aromatic nucleophilic substitution reaction to cause the 2, 4-dinitrofluorobenzene unit to be separated from the hydrogen sulfide fluorescent probe (formula 4), so that a molecular fluorescent signal is recovered, and the detection of hydrogen sulfide is realized. Through tests, the detection sensitivity of the hydrogen sulfide detection fluorescent probe is up to 36 nanomole, and the hydrogen sulfide detection fluorescent probe can be used for monitoring hydrogen sulfide in a special environment of cells.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification, illustrate exemplary embodiments of the invention and together with the description serve to explain the invention and not to limit the invention.
FIG. 1 is a nuclear magnetic resonance hydrogen spectrum of a viscosity and polarity responsive plateau fluorescent probe molecule synthesized according to a first embodiment of the present invention.
FIG. 2 is an enlarged view of the NMR spectrum of the first embodiment of the synthesized viscosity and polarity responsive plateau fluorescent probe molecule in the region of 6.0-8.8 chemical shifts.
FIG. 3 is the NMR spectrum of a fluorescent probe for hydrogen sulfide detection synthesized in the third embodiment of the present invention.
FIG. 4 is a graph of fluorescence emission of the viscosity and polarity response type platform fluorescent probe molecules synthesized according to the first embodiment of the present invention in a system with different viscosities of glycerol-water composition.
FIG. 5 is a graph showing the relationship between fluorescence intensity and viscosity in different viscosity systems of glycerol-water compositions for the viscosity and polarity response type platform fluorescent probe molecules synthesized according to the first embodiment of the present invention.
FIG. 6 is a graph showing fluorescence emission of the viscosity and polarity responsive plateau fluorescent probe molecule synthesized according to the first embodiment of the present invention in different polar solvents.
FIG. 7 is a graph showing the quantitative relationship between the fluorescence emission wavelength of the viscosity and polarity response type platform fluorescent probe molecule synthesized in the first embodiment of the present invention in different polarity solvents.
FIG. 8 is a fluorescence spectrum of the hydrogen sulfide detecting fluorescent probe synthesized according to the second embodiment of the present invention after reacting with 28 analytes, respectively.
FIG. 9 is a graph showing the fluorescence intensity at 530nm after the hydrogen sulfide detecting fluorescent probe synthesized according to the second embodiment of the present invention reacts with 28 analytes, respectively.
FIG. 10 is a graph showing fluorescence intensities at 530nm after reaction with sodium sulfide in the presence of various interferents, in the hydrogen sulfide detecting fluorescent probe synthesized according to the second embodiment of the present invention.
FIG. 11 is a graph showing fluorescence intensities at different time points of the reaction between the hydrogen sulfide-detecting fluorescent probe synthesized in the second embodiment of the present invention and sodium sulfide.
FIG. 12 is a graph showing the fluorescence intensity of the hydrogen sulfide detecting fluorescent probe synthesized in the second example of the present invention after reacting with hydrogen sulfide at different concentrations for the same time.
FIG. 13 is a linear relationship graph of fluorescence intensity and hydrogen sulfide concentration after the hydrogen sulfide detecting fluorescent probe synthesized in the second embodiment of the present invention reacts with hydrogen sulfide for the same time.
FIG. 14 is a graph showing the results of the cell viability of Hela cells treated with different concentrations of the hydrogen sulfide detecting fluorescent probe synthesized in the second example.
FIG. 15 is a graph showing the results of the fluorescence confocal imaging application of the fluorescent probe for detecting hydrogen sulfide synthesized in the second embodiment of the present invention to hydrogen sulfide in cells.
Detailed Description
In the following description, further 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 particular 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. Unless otherwise indicated, the drugs or agents used in the present invention are used according to the instructions of the product or by the conventional methods in the art. The process of the present 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 response type platform fluorescent probe, referring to reaction scheme 1, comprises the following steps:
(1) 4mmol of 4-diethylamino salicylaldehyde (formula 2), 1mmol of the compound represented by formula 1 and 20mL of methanol were weighed in a three-necked flask, and the system was heated at 50 ℃ for reaction.
(2) Monitoring the reaction by TLC, 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) carrying out suction filtration on the solid-liquid mixture to obtain an orange solid, namely the viscosity and polarity response type platform fluorescent probe, as shown in the formula 3.
The nmr hydrogen spectra of the viscosity and polarity responsive plateau fluorescent probe of formula 3 synthesized in this example are shown in fig. 1 and 2. Wherein, FIG. 1 is the nuclear magnetic resonance hydrogen spectrum of the fluorescent probe molecule, and FIG. 2 is the enlarged view of the nuclear magnetic resonance hydrogen spectrum of the fluorescent probe molecule in the region with the chemical shift of 6.0-8.8. The structural formula 3 of the viscosity and polarity response type platform fluorescent probe can be determined from fig. 1 and 2.
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 method for preparing a viscosity and polarity response type platform fluorescent probe, which is referred to the reaction scheme 1, comprises the following steps:
(1) 1mmol of 4-diethylamino salicylaldehyde (formula 2), 1mmol of the compound represented by formula 1 and 5mL of DMF were weighed into a three-necked flask, and the system was heated at 100 ℃ for reaction.
(2) Monitoring the reaction by TLC, and cooling the reaction liquid to room temperature when the reaction intermediate disappears; slowly adding the reaction solution into 50mL of saturated saline solution, and precipitating a large amount of solid; and (3) carrying out suction filtration on the solid-liquid mixture to obtain an orange solid, namely the viscosity and polarity response type platform fluorescent probe, as shown in the 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 preparation method of a hydrogen sulfide detection fluorescent probe refers to reaction scheme 2 and comprises the following steps:
(1) weighing 0.5mmol of the viscosity and polarity response type platform fluorescent probe synthesized in the first embodiment and shown in formula 3, 0.6mmol of 2, 4-dinitrofluorobenzene, 0.55mmol of triethylamine and 10mL of dichloromethane in a three-neck flask; the mixture was heated at 35 ℃ for reaction.
(2) And monitoring the reaction completion by TLC, cooling the reaction solution to room temperature, extracting and washing the reaction solution with deionized water for three times, drying an organic layer with anhydrous sodium sulfate, and removing the solvent by rotary evaporation to obtain a crude product. And 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, as shown in a formula 4.
The nmr hydrogen spectrum of the hydrogen sulfide detecting fluorescent probe of formula 4 synthesized in this example is shown in fig. 3, and it can be determined from fig. 3 that the nmr hydrogen spectrum of the hydrogen sulfide detecting fluorescent probe is shown in 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 ＝2.50 Hz,1H),6.25(d,J＝2.50Hz,1H),4.43(q,J＝7.10Hz,2H),3.49(q,J＝7.05 Hz,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 refers to the reaction scheme 2, and comprises the following steps:
(1) weighing 0.5mmol of the viscosity and polarity response type platform fluorescent probe synthesized in the formula 3 synthesized in the first embodiment, 0.6mmol of 2, 4-dinitrofluorobenzene, 0.55mmol of triethylamine and 10mL of acetonitrile in a three-neck flask; the mixture was heated at 35 ℃ for reaction.
(2) TLC to monitor the reaction is complete, stop the reaction, remove the solvent by rotary evaporation, dissolve the product with 10mL dichloromethane, wash with saturated brine three times, dry the organic layer with anhydrous sodium sulfate, and remove the solvent by rotary evaporation to obtain the crude product. And 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, as shown in a formula 4.
The structural characterization data of the viscosity and polarity response type 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 ＝2.50 Hz,1H),6.25(d,J＝2.50Hz,1H),4.43(q,J＝7.10Hz,2H),3.49(q,J＝7.05 Hz,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 preparation method of a hydrogen sulfide detection fluorescent probe refers to reaction scheme 3 and comprises the following steps:
(1) weighing 0.5mmol of the viscosity and polarity response type platform fluorescent probe synthesized in the first embodiment and shown in formula 3, 0.5mmol of 2, 4-dinitrofluorobenzene, 0.5mmol of potassium carbonate and 10mL of dichloromethane in a three-neck flask; the mixture was heated at 35 ℃ for reaction.
(2) TLC monitors the reaction is complete, the reaction liquid is cooled to room temperature, the reaction liquid is extracted and washed by deionized water for three times, an organic layer is dried by anhydrous sodium sulfate, and the solvent is removed by rotary evaporation to obtain a crude product. And 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 preparation method of a hydrogen sulfide detection fluorescent probe refers to a reaction route 4 and comprises the following steps:
(1) weighing 0.6mmol of the viscosity and polarity response type platform fluorescent probe synthesized in the first embodiment and shown in formula 3, 0.9mmol of 2, 4-dinitrofluorobenzene, 0.9mmol of triethylamine and 10mL of chloroform in a three-neck flask; the mixture was heated at 35 ℃ for the reaction.
(2) And monitoring the reaction completion by TLC, cooling the reaction solution to room temperature, extracting and washing the reaction solution with deionized water for three times, drying an organic layer with anhydrous sodium sulfate, and removing the solvent by rotary evaporation to obtain a crude product. And 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 testing
1. Weighing the viscosity and polarity response type platform fluorescent probe molecules (formula 3) synthesized in the first embodiment, dissolving the platform fluorescent probe molecules in DMSO to prepare a 1mM probe molecule solution; the 1mM probe molecule solution is prepared into 10 mu M/L probe solution in different viscosity systems consisting of glycerol and water.
(1) The change in fluorescence intensity in the probe solution was measured, and the result is shown in FIG. 4. As can be seen from this FIG. 4, the viscosity and polarity responsive platform fluorescent probe molecules exhibit unique sensitivity to different viscosities of solution systems, with the fluorescence intensity increasing with increasing viscosity of the system.
(2) Testing the saidThe fluorescence intensity of the probe solution was linearly fitted to the viscosity of the system, and the results are shown in FIG. 5. As can be seen from FIG. 5, the fluorescence intensity of the viscosity and polar response type platform fluorescent probe molecule at 530nm has a good linear relationship with the viscosity of the system (R) 2 0.99), can be used as an excellent viscosity change fluorescent probe.
2. Weighing the viscosity and polarity response type platform fluorescent probe molecules (formula 3) synthesized in the first embodiment, dissolving the molecules in DMSO to prepare a 1mM probe molecule solution; taking the above 1mM probe molecule solution in different polar solvents (including ethyl acetate (EtOAc), methanol (MeOH), ethanol (EtOH), Isopropanol (IPA), dimethyl sulfoxide (DMSO), and water (H) 2 O)), 10. mu.M/L of a probe solution in a solvent of different polarity was prepared. The fluorescence emission of the probe solutions of different polar solvents was separately tested, 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. Weighing the viscosity and polarity response type platform fluorescent probe molecules (formula 3) synthesized in the first embodiment, dissolving the platform fluorescent probe molecules in DMSO to prepare a 1mM probe molecule solution; taking the above 1mM probe molecule solution in different polar solvents (including ethyl acetate (EtOAc), methanol (MeOH), ethanol (EtOH), Isopropanol (IPA), dimethyl sulfoxide (DMSO), and water (H) 2 O)) was prepared as a 10. mu.M/L probe solution in a solvent of different polarity.
(1) The probe solutions of different polarity solvents were tested separately for their quantitative relationship of fluorescence emission wavelength, and the results are shown in fig. 7. As can be seen from FIG. 7, the maximum emission wavelength of the viscosity and polarity response type platform fluorescent probe molecule in these solvents is in quantitative relation to the dielectric constant (R) 2 0.99), indicating that the probe molecule can be used for quantitative analysis of environmental polarity changes.
4. The hydrogen sulfide detection fluorescent probe (formula 4) synthesized in the third example was weighed and dissolved in DMSO to prepare a 1mM probe molecule solution. Taking the probe molecule solution in Tirs-HCl buffer solution (DMSO/H) 2 O1/1, v/v, pH 7.4), prepared as 10 μ M/L probeAnd (4) a buffer solution.
To three identical aliquots of the probe buffer were added analytes: cation and anion (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 anion), amino acids (glycine, glutamic acid, arginine, lysine, tyrosine, aspartic acid, histidine), biological thiols (cysteine, glutamic acid), to give three analyte solutions each at a concentration of 200 μ M/L.
The fluorescence spectra and fluorescence intensities of the above three analyte solutions were measured, and the results are shown in fig. 8 and 9. As can be seen from fig. 8 and 9, the fluorescence intensity of the analyte solution in the hydrogen sulfide atmosphere changes significantly, and the fluorescence emission peak is about 530nm, but no significant fluorescence change is observed in the other analyte atmospheres. The above experiment confirms 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 (formula 4) synthesized in the third example was weighed and dissolved in DMSO to prepare a 1mM probe molecule solution. Taking the probe molecule solution in Tirs-HCl buffer solution (DMSO/H) 2 O-1/1, v/v, pH 7.4), prepared as 10 μ M/L probe buffer.
To three identical aliquots of the probe buffer were added analytes: cation and anion (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 anion), amino acids (glycine, glutamic acid, arginine, lysine, tyrosine, aspartic acid, histidine), biological thiols (cysteine, glutamic acid), to give three analyte solutions each at a concentration of 200 μ M/L.
To each of the three analyte solutions was added 200. mu.M/L sodium sulfide. The three analyte solutions (containing sodium sulfide) obtained were tested for fluorescence intensity at 530nm before and after the reaction, and the results are shown in FIG. 10. As can be seen from the figure, the hydrogen sulfide detection fluorescent probe (formula 4) does not show significant fluorescence change in the atmosphere of other interferents, and the fluorescence intensity is obviously changed after hydrogen sulfide is added into the interferents, which indicates that the probe molecule can be used for specific recognition of hydrogen sulfide molecules.
6. The hydrogen sulfide detection fluorescent probe (formula 4) synthesized in the third example was weighed and dissolved in DMSO to prepare a 1mM probe molecule solution. Taking the probe molecule solution in Tirs-HCl buffer solution (DMSO/H) 2 O-1/1, v/v, pH 7.4), prepared as 10 μ M/L probe buffer.
To the probe buffer, 200. mu.M/L sodium sulfide was added and the fluorescence intensity was measured at different time points of the reaction, and the results are shown in FIG. 11. From the figure, the emission at 530nm almost reaches saturation within 10min, and the fluorescence is enhanced by 95 times, which shows that the probe molecule has a rapid response to hydrogen sulfide.
7. The hydrogen sulfide detection fluorescent probe (formula 4) synthesized in the third example was weighed and dissolved in DMSO to prepare a 1mM probe molecule solution. Taking the probe molecule solution in Tirs-HCl buffer solution (DMSO/H) 2 O-1/1, v/v, pH 7.4), prepared as 10 μ M/L probe buffer.
Sodium sulfide was added to the probe buffer at different 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, with the results shown in fig. 12. As can be seen from this figure, the fluorescence intensity of the probe solution gradually increased with the 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 hydrogen sulfide concentration, and the results are shown in FIG. 13. As can be seen from the graph, in the concentration range of 0-20 μm, there is a linear relationship between the fluorescence intensity of the probe solution and the concentration of hydrogen sulfide, and the probe solution can be used for quantitative detection of hydrogen sulfide.
8. The hydrogen sulfide detection fluorescent probe (formula 4) synthesized in the third example is weighed and dissolved in DMSO to prepare a probe molecule mother solution.
Adopting a DMEM culture medium to culture Hela cells; the probe molecule mother liquor is used as mother liquor, and the MTT method is used for testing the survival rate of Hela cells after the probe molecule mother liquor with different concentrations is treated, and the result is shown in FIG. 14. As can be seen from the figure, the hydrogen sulfide detection fluorescent probe (formula 4) is almost non-toxic to cells at a concentration of 0-100 μ M, and has a high cell survival rate.
9. HeLa cells were cultured in DMEM medium for 12 hours using the hydrogen sulfide-detecting fluorescent probe (formula 4) synthesized in the third example, and then the cells were incubated in DMEM medium containing 200. mu.M sodium sulfide for 12 hours, and fluorescence imaging was performed on the cells on a fluorescence confocal microscope, the results of which are shown in FIG. 15. The figure shows that the fluorescent probe has higher signal-to-noise ratio to the hydrogen sulfide in the cells, and can be used as a high-contrast imaging probe to detect the exogenous hydrogen sulfide in the living cells.
The above description is only illustrative of several embodiments of the present invention and should not be taken as limiting the scope of the invention. It should be noted that other persons skilled in the art can make modifications, substitutions, improvements and the like without departing from the spirit and scope of the present invention, and all of them belong to the protection scope of the present invention. Therefore, the scope of the invention should be determined from the description and claims.
Claims (10)
1. A viscosity and polarity response type platform fluorescent probe has a structure shown in a formula 3;
in said formula 3, the substituent R 1 Is a hydrocarbyl group, preferably methyl or ethyl; substitutionRadical R 2 is-CN, -CONH 2 and-COOH.
2. The process for synthesizing a viscosity and polarity response type platform fluorescent probe according to claim 1, comprising the steps of: taking a compound shown in a formula 1 and 4-diethylamino salicylaldehyde as raw materials, heating to react, and separating out a target product shown in a formula 3 to obtain the compound;
in the formula 1, a substituent R 1 Is a hydrocarbyl group, preferably methyl or ethyl; substituent R 2 is-CN, -CONH 2 and-COOH.
3. The process for synthesizing the viscosity and polarity response type platform fluorescent probe according to claim 2, wherein the molar ratio of the compound represented by formula 1 to 4-diethylamino salicylaldehyde is in the range of 1: 1-4: 1.
4. the process for synthesizing the viscosity and polarity response type platform fluorescent probe according to claim 2 or 3, wherein the compound represented by formula 1 and 4-diethylamino salicylaldehyde are dissolved in a solvent and then heated for reaction;
preferably, the solvent comprises: any one of methanol, ethanol, N-dimethylformamide and dimethyl sulfoxide;
preferably, the temperature of the heating reaction is 50-100 ℃.
5. A hydrogen sulfide detection fluorescent probe has a structure shown in a formula 4;
in said formula 4, the substituent R 1 Is a hydrocarbyl group, preferably methyl or ethyl; substitutionRadical R 2 is-CN, -CONH 2 and-COOH.
6. The process for synthesizing the hydrogen sulfide detection fluorescent probe according to claim 5, which 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, reacting under the catalysis of alkali, and separating out a target product shown in a formula 4 to obtain the fluorescent probe.
7. The process for synthesizing the hydrogen sulfide detection fluorescent probe according to claim 6, wherein the molar ratio of the viscosity and polarity response type platform fluorescent probe shown in formula 3 to 2, 4-dinitrofluorobenzene is in the range of 1: 1-1: 1.5.
8. the synthesis process of the hydrogen sulfide detection fluorescent probe according to claim 6, wherein the viscosity and polarity response type platform fluorescent probe shown in formula 3 and 2, 4-dinitrofluorobenzene are dissolved in a solvent and then react; preferably, the solvent comprises: any one of dichloromethane, chloroform and acetonitrile.
9. The process for synthesizing a hydrogen sulfide detection fluorescent probe according to claim 6, wherein the addition ratio of the base is one time or more of the molar weight of the raw material represented by formula 3; preferably, the base comprises any one of triethylamine and potassium carbonate.
10. Use of the viscosity and polarity responsive platform fluorescent probe of claim 1 or the hydrogen sulfide detecting fluorescent probe of claim 5 in the biological and medical field, preferably for in vivo monitoring of hydrogen sulfide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210483797.XA CN114805262B (en) | 2022-05-05 | 2022-05-05 | Viscosity and polarity response type platform fluorescent probe, hydrogen sulfide detection fluorescent probe, and synthesis process and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210483797.XA CN114805262B (en) | 2022-05-05 | 2022-05-05 | Viscosity and polarity response type platform fluorescent probe, hydrogen sulfide detection fluorescent probe, and synthesis process and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114805262A true CN114805262A (en) | 2022-07-29 |
| CN114805262B CN114805262B (en) | 2024-03-26 |
Family
ID=82512318
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210483797.XA Active CN114805262B (en) | 2022-05-05 | 2022-05-05 | Viscosity and polarity response type platform fluorescent probe, hydrogen sulfide detection fluorescent probe, and synthesis process and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114805262B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2024054449A1 (en) * | 2022-09-09 | 2024-03-14 | Saudi Arabian Oil Company | Aqueous fluid compositions and barite scale removal therewith |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104419401A (en) * | 2013-08-28 | 2015-03-18 | 苏州罗兰生物科技有限公司 | Fluorescent probe for detecting hydrogen sulfide by virtue of fluorescence enhancement as well as synthetic method and application of fluorescent probe |
| 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 |
| 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 |
-
2022
- 2022-05-05 CN CN202210483797.XA patent/CN114805262B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104419401A (en) * | 2013-08-28 | 2015-03-18 | 苏州罗兰生物科技有限公司 | Fluorescent probe for detecting hydrogen sulfide by virtue of fluorescence enhancement as well as synthetic method and application of fluorescent probe |
| 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 |
| 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 |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2024054449A1 (en) * | 2022-09-09 | 2024-03-14 | Saudi Arabian Oil Company | Aqueous fluid compositions and barite scale removal therewith |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114805262B (en) | 2024-03-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Liu et al. | A novel near-infrared fluorescent probe with a large Stokes shift for biothiol detection and application in in vitro and in vivo fluorescence imaging | |
| CN105712964B (en) | Preparation method and application of thiol fluorescent probe based on coumaroyl hydrazide | |
| CN111253935B (en) | Two-photon fluorescent probe for detecting polarity and viscosity through two channels and preparation method and application thereof | |
| CN107298654A (en) | The fluorescence probe of hydrogen sulfide in a kind of detection endoplasmic reticulum | |
| CN110511203B (en) | Mustard gas fluorescent probe and preparation and application thereof | |
| CN110698454B (en) | Isophorone hydrogen sulfide fluorescent probe and preparation method and application thereof | |
| CN109867611A (en) | A kind of for red wine and in vivo water-soluble two-photon hydrogen sulfide fluorescence probe and its preparation method and application of sulfurated hydrogen detection | |
| CN105524055A (en) | Preparation and application of fluorescent probe capable of being used for distinguishing cysteine/homocysteine and glutathione | |
| CN109535147A (en) | A kind of formaldehyde fluorescence probe of quick response and its preparation method and application | |
| CN111892923B (en) | Two-photon fluorescence viscosity probe based on dinitrile vinyl group and preparation method and application thereof | |
| WO2017078623A9 (en) | Background-free fluorescent probes for live cell imaging | |
| Qi et al. | Near-infrared turn-on fluorescent probe for discriminative detection of Cys and application in in vivo imaging | |
| CN112500386A (en) | Near-infrared HClO fluorescent probe based on pyrrazone oxime, and preparation and application thereof | |
| CN114805262B (en) | Viscosity and polarity response type platform fluorescent probe, hydrogen sulfide detection fluorescent probe, and synthesis process and application thereof | |
| CN110643355A (en) | Fluorescent probe for detecting polarity of endoplasmic reticulum as well as preparation method and application thereof | |
| CN109438426B (en) | Half-cyanine-based reactive type Hg2+ fluorescent probe Cy-PT and preparation method and application thereof | |
| CN110092773A (en) | A kind of oxa anthracenes derivative and its preparation method and application | |
| Rong et al. | A naphthalimide-indole fused chromophore-based fluorescent probe for the detection of biothiol with red emission and a large Stokes shift | |
| CN113999159B (en) | Viscosity-sensitive fluorescent probe, and preparation method and application thereof | |
| CN113582985B (en) | Mitochondrion targeted pH and viscosity dual-channel detection fluorescent probe and preparation method and application thereof | |
| Tang et al. | A new metal-free near-infrared fluorescent probe based on nitrofuran for the detection and bioimaging of carbon monoxide releasing molecule-2 in vivo | |
| CN114230494A (en) | Synthesis of large Stokes shift near-infrared fluorescent probe and application thereof in detecting hydrogen sulfide | |
| CN110229203B (en) | Hexosamine fluorescent probe and preparation method and application thereof | |
| CN111925316B (en) | Two-photon fluorescence polar probe based on 4-fluorophenylethynyl and preparation method and application thereof | |
| CN111333649B (en) | SNAP-tag technology-based cell membrane fluorescent probe and preparation and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |