CN107033177A - A kind of hypersensitive high selection peroxynitrite colorimetric ratio fluorescent probe using pinacol borate as identification receptor - Google Patents
A kind of hypersensitive high selection peroxynitrite colorimetric ratio fluorescent probe using pinacol borate as identification receptor Download PDFInfo
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- CN107033177A CN107033177A CN201710218670.4A CN201710218670A CN107033177A CN 107033177 A CN107033177 A CN 107033177A CN 201710218670 A CN201710218670 A CN 201710218670A CN 107033177 A CN107033177 A CN 107033177A
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- peroxynitrite
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- CMFNMSMUKZHDEY-UHFFFAOYSA-N peroxynitrous acid Chemical compound OON=O CMFNMSMUKZHDEY-UHFFFAOYSA-N 0.000 title claims abstract description 53
- 239000007850 fluorescent dye Substances 0.000 title abstract description 8
- BMQDAIUNAGXSKR-UHFFFAOYSA-N (3-hydroxy-2,3-dimethylbutan-2-yl)oxyboronic acid Chemical compound CC(C)(O)C(C)(C)OB(O)O BMQDAIUNAGXSKR-UHFFFAOYSA-N 0.000 title abstract description 7
- 206010020751 Hypersensitivity Diseases 0.000 title abstract description 4
- -1 rhodamine compound Chemical class 0.000 claims abstract description 16
- 150000001875 compounds Chemical class 0.000 claims description 8
- 238000002360 preparation method Methods 0.000 claims description 6
- 125000003545 alkoxy group Chemical group 0.000 claims description 4
- 125000000217 alkyl group Chemical group 0.000 claims description 4
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 4
- 125000004185 ester group Chemical group 0.000 claims description 4
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 4
- GQPLMRYTRLFLPF-UHFFFAOYSA-N nitrous oxide Inorganic materials [O-][N+]#N GQPLMRYTRLFLPF-UHFFFAOYSA-N 0.000 claims description 2
- 229910019142 PO4 Inorganic materials 0.000 claims 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims 1
- 239000010452 phosphate Substances 0.000 claims 1
- 239000000523 sample Substances 0.000 abstract description 62
- 238000001514 detection method Methods 0.000 abstract description 17
- 238000000034 method Methods 0.000 abstract description 7
- 230000000694 effects Effects 0.000 abstract description 2
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 18
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical group CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 14
- 239000000047 product Substances 0.000 description 13
- 238000012360 testing method Methods 0.000 description 12
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 10
- 238000002189 fluorescence spectrum Methods 0.000 description 9
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 8
- BMIBJCFFZPYJHF-UHFFFAOYSA-N 2-methoxy-5-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine Chemical compound COC1=NC=C(C)C=C1B1OC(C)(C)C(C)(C)O1 BMIBJCFFZPYJHF-UHFFFAOYSA-N 0.000 description 7
- 239000002253 acid Substances 0.000 description 7
- 229960004194 lidocaine Drugs 0.000 description 7
- ALTGURJQVWBILJ-UHFFFAOYSA-N n-(dimethylamino)formamide Chemical compound CN(C)NC=O ALTGURJQVWBILJ-UHFFFAOYSA-N 0.000 description 7
- SVENKGKXRLHCTC-UHFFFAOYSA-N B(O)(O)O.BrCC=1C=CC=CC1 Chemical class B(O)(O)O.BrCC=1C=CC=CC1 SVENKGKXRLHCTC-UHFFFAOYSA-N 0.000 description 6
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 6
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 6
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 description 6
- FJDQFPXHSGXQBY-UHFFFAOYSA-L caesium carbonate Chemical compound [Cs+].[Cs+].[O-]C([O-])=O FJDQFPXHSGXQBY-UHFFFAOYSA-L 0.000 description 6
- 239000012043 crude product Substances 0.000 description 6
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 6
- XXQBEVHPUKOQEO-UHFFFAOYSA-N potassium superoxide Chemical compound [K+].[K+].[O-][O-] XXQBEVHPUKOQEO-UHFFFAOYSA-N 0.000 description 6
- 238000000926 separation method Methods 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 125000002147 dimethylamino group Chemical group [H]C([H])([H])N(*)C([H])([H])[H] 0.000 description 5
- 238000000605 extraction Methods 0.000 description 5
- 150000002500 ions Chemical class 0.000 description 5
- 230000035945 sensitivity Effects 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 238000001228 spectrum Methods 0.000 description 5
- 238000003786 synthesis reaction Methods 0.000 description 5
- BVKZGUZCCUSVTD-UHFFFAOYSA-M Bicarbonate Chemical compound OC([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-M 0.000 description 4
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 4
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 4
- 229910002651 NO3 Inorganic materials 0.000 description 4
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 4
- FKNQFGJONOIPTF-UHFFFAOYSA-N Sodium cation Chemical compound [Na+] FKNQFGJONOIPTF-UHFFFAOYSA-N 0.000 description 4
- PTFCDOFLOPIGGS-UHFFFAOYSA-N Zinc dication Chemical compound [Zn+2] PTFCDOFLOPIGGS-UHFFFAOYSA-N 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 229910001425 magnesium ion Inorganic materials 0.000 description 4
- 150000002978 peroxides Chemical class 0.000 description 4
- 229910001415 sodium ion Inorganic materials 0.000 description 4
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 3
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 3
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical group [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 description 3
- 238000000862 absorption spectrum Methods 0.000 description 3
- 229940006460 bromide ion Drugs 0.000 description 3
- 229910001424 calcium ion Inorganic materials 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 229960003180 glutathione Drugs 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 3
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 2
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 2
- NPYPAHLBTDXSSS-UHFFFAOYSA-N Potassium ion Chemical compound [K+] NPYPAHLBTDXSSS-UHFFFAOYSA-N 0.000 description 2
- REDXJYDRNCIFBQ-UHFFFAOYSA-N aluminium(3+) Chemical compound [Al+3] REDXJYDRNCIFBQ-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 229910001431 copper ion Inorganic materials 0.000 description 2
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 2
- 235000018417 cysteine Nutrition 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 229940005654 nitrite ion Drugs 0.000 description 2
- 229910001414 potassium ion Inorganic materials 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical group [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 2
- CIHOLLKRGTVIJN-UHFFFAOYSA-N tert‐butyl hydroperoxide Chemical compound CC(C)(C)OO CIHOLLKRGTVIJN-UHFFFAOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- BZSXEZOLBIJVQK-UHFFFAOYSA-N 2-methylsulfonylbenzoic acid Chemical compound CS(=O)(=O)C1=CC=CC=C1C(O)=O BZSXEZOLBIJVQK-UHFFFAOYSA-N 0.000 description 1
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- KWGACYZYFZTYRN-UHFFFAOYSA-N OC(C)(C)C(C)(C)O.B(O)(O)O.BrCC1=CC=CC=C1 Chemical class OC(C)(C)C(C)(C)O.B(O)(O)O.BrCC1=CC=CC=C1 KWGACYZYFZTYRN-UHFFFAOYSA-N 0.000 description 1
- 239000005708 Sodium hypochlorite Substances 0.000 description 1
- 241001104043 Syringa Species 0.000 description 1
- 235000004338 Syringa vulgaris Nutrition 0.000 description 1
- WITZXUPUMRPBJS-UHFFFAOYSA-N [B].BrCC1=CC=CC=C1 Chemical compound [B].BrCC1=CC=CC=C1 WITZXUPUMRPBJS-UHFFFAOYSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 238000003968 anodic stripping voltammetry Methods 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- 229910000024 caesium carbonate Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000002848 electrochemical method Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229910001448 ferrous ion Inorganic materials 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 238000004313 potentiometry Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic Table
- C07F5/02—Boron compounds
- C07F5/04—Esters of boric acids
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- 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"
- G01N21/643—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" non-biological material
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- 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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- 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
- C09K2211/1096—Heterocyclic compounds characterised by ligands containing other heteroatoms
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- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
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- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Optics & Photonics (AREA)
- Chemical Kinetics & Catalysis (AREA)
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- General Health & Medical Sciences (AREA)
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- Engineering & Computer Science (AREA)
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- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The present invention relates to a kind of hypersensitive high selection peroxynitrite colorimetric ratio fluorescent probe using pinacol borate as identification receptor.Specifically, probe of the invention is rhodamine compound of the class using pinacol borate as identification receptor, and it can be used for the detection of peroxynitrite as peroxynitrite fluorescence probe.This kind of probe can realize at least one in following technique effect：Hypersensitive recognizes peroxynitrite with high selectivity；Quickly peroxynitrite can be realized and responded；The instant detection to peroxynitrite can be realized；Property is stable, can for a long time preserve and use；Be conducive to detecting peroxynitrite in actual sample.
Description
Technical field
The present invention relates to pinacol borate peroxynitrite is used as the rhodamine compound of identification receptor
Colorimetric ratio fluorescent probe, it is applied in being recognized under given conditions to peroxynitrite quick high-selectivity, or
It can in determination sample peroxynitrite concentration.
Background technology
Peroxynitrite is the active oxidizer generated by nitric oxide and peroxide reactions, is disappeared with sterilization
The effect of poison, plays an important role in human immune system.If peroxynitrite abnormal accumulation, itself and two grades of generations
Thanking to thing can react with protein-based, lipid and nucleic acid material, and then promote cytotoxic effect to cause cell death and rush
Enter inflammatory reaction, the other illnesss such as Alzheimer disease, arthritis, autoimmune disease can be caused so as to human health
Constitute grave danger.Therefore, the concentration of detection organism endoperoxides nitrite has become an important problem.
Currently, the detection means of common peroxynitrite be mainly electrochemical method (such as anodic stripping voltammetry,
Redox potentiometry etc.).But this means are not only expensive but also cumbersome in actual applications, and usually need special laboratory apparatus
And high professional qualification professional is operated.Therefore, efficient, cheap, simple and direct peroxynitrite detection means turns into weight
The research topic wanted.
In the detection method of various ion/moleculars, colorimetric fluorescence probe detection method can be entered at the scene due to simple to operate
Exercise and use, selectivity and sensitivity are high, detection range is wide, the response time is quick, detection process sample is not destroyed and extremely
It is noticeable.But the fluorescence probe reported at present still suffers from some problems, including sensitivity is undesirable, poor selectivity, pH are steady
Qualitative difference and synthesis complexity etc..In a word, development high sensitivity high selectivity, the simple colorimetric fluorescence probe of synthesis are this areas
Technical staff's urgent problem.
The content of the invention
It is glimmering that a kind of colorimetric for preparing simple fast high-sensitive high selectivity identification peroxynitrite is badly in need of in this area
Light probe, so as to effective detection peroxynitrite.Therefore, the present invention has synthesized the novel peroxinitrites of a class
The colorimetric fluorescence probe of salt, its synthesis step is simple, sensitivity is high, stability is high, and/or selectivity high, and/or can be quick
Recognize peroxynitrite.
Specifically, the invention provides a kind of peroxynitrite colorimetric fluorescence probe, it is rhodamine chemical combination
Thing, using pinacol borate as specific recognition acceptor, its structure is as follows：
It is preferred that, fluorescence probe of the invention is：
In the preparation method of the peroxynitrite colorimetric fluorescence probe of the present invention, reaction temperature is 25-120 DEG C；
Reaction time is 2-24h；And/or rhodamine compound and the mol ratio of 4- bromomethyl benzene boric acid pinacol esters are about 1：1 to
1：2, preferably 1：1.5 or 1：1.6.
Present invention also offers the detection system for detecting peroxynitrite concentration in sample (such as river sample)
Agent or kit, it includes the probe of the present invention.Preferably, detection preparation of the invention or kit also include the use of product
Specification.It is further preferred that the kit of the present invention also includes the buffering for being used for determining the peroxynitrite concentration in sample
Agent.
Present invention also offers the method for peroxynitrite concentration in detection sample (such as river sample), it includes
The step of probe of the present invention is contacted with sample to be tested.
Prepared present invention also offers the probe of the present invention for detecting that peroxidating is sub- in sample (such as river sample)
Purposes in the preparation of nitrate concentration.
Prepared present invention also offers the probe of the present invention for detecting that peroxidating is sub- in sample (such as river sample)
Purposes in the kit of nitrate concentration.
The peroxynitrite fluorescence probe of the present invention can be acted on peroxynitrite, produce fluorescence spectrum
Change, so as to realize the quantitative detection to peroxynitrite
Specifically, peroxynitrite fluorescence probe of the invention respectively with hydrogen peroxide, hypochlorous acid, bromide ion with
And other ions are acted on can not cause the obvious change of fluorescence spectrum, so as to realize the selection to peroxynitrite
Property identification.
Selectively, the stability of peroxynitrite fluorescence probe of the invention is good, and then can for a long time preserve and make
With.
Further, peroxynitrite fluorescence probe of the invention is quick high-selectivity peroxynitrite ratio
Color ratio probes, and synthesis is simple, is conducive to commercialized popularization and application.
Brief description of the drawings
Fig. 1 is influence and quantitative line of the various concentrations peroxynitrite (0-8 μM) to probe (5 μM) absorption spectrum
Sexual intercourse.
Fig. 2 is various concentrations Hg2+The influence and linear quantitative relation of (0-2 μM) to probe (5 μM) fluorescence spectrum.
Fig. 3 is test result of the probe (5 μM) to peroxynitrite (5 μM) response time.
Fig. 4 is influence of the different analytes to probe (5 μM) fluorescence spectrum.Wherein：1. blank；2. potassium ion K+；3. calcium
Ion Ca2+；4. sodium ion Na+；5. magnesium ion Mg2+；6. aluminium ion Al3+；7. zinc ion Zn2+；8. iron ion Fe3+；9. it is ferrous from
Sub- Fe2+；10. copper ion Cu2+；11. nitrate ion NO3 -；12. nitrite ion NO2 -；13. cysteine CYS；(3mM)
14. glutathione GSH (3mM)；15. hydrogen peroxide H2O2(200μM)；16. tertbutanol peroxide TBHP；17. hydroxyl radical free radical
OH；18. tert-Butanol peroxide free radical OtBu；19. nitric oxide NO；20. singlet oxygen1O2；21. potassium superoxide KO2；22. time
Sodium chlorate NaClO；23. carbonate CO3 2-；24. bisulfite HSO3 -；25. bicarbonate radical HCO3 -；26. bromide ion Br-；27.
Sulphion S2-；28. peroxynitrite ONOO-。
Fig. 5 is open hole detection of the probe to peroxynitrite, and wherein concentration and probe concentration is 20 μM, peroxynitrite
Concentration is respectively 0,10,15,20 μM, test system is ethanol：Water (1:9, v/v), PBS (20Mm, pH=7.4) test temperature is
25℃。
Embodiment：
The invention provides the synthetic route of above-mentioned quick high-selectivity peroxynitrite fluorescence probe, method and its
Spectrum property.
The peroxynitrite fluorescence probe of the present invention is rhodamine of the class using pinacol borate as identification receptor
Class compound, it has following general structure
In above formula：R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12, R13, R14, R15And R16For hydrogen atom, straight chain or
Branched alkyl, straight or branched alkoxyl, sulfonic group, ester group, carboxyl；R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12,
R13, R14, R15And R16Can be with identical or different.
The synthetic route and method of such peroxynitrite colorimetric ratio fluorescent probe are as follows：
Specifically, fluorescence probe of the invention can be prepared via a method which, by certain mol proportion (such as 1：1-1：
2) rhodamine compound (such as synthetic product of 1,6- dihydroxy naphthlenes and 4- lignocaine ketone acids) and 4- bromomethyl benzene boron
Sour pinacol ester is dissolved in dimethylamino formamide (DMF), adds cesium carbonate and KI, when then stirring one section at room temperature
Between (such as 24h), then extract, rotate obtain crude product.Use dichloromethane：Methanol (30:1, v/v) system carries out column chromatography
Separation, obtains lightpink pure product 588.6mg, and yield is 90 ﹪.
Therefore, prepared present invention also offers 4- bromomethyl benzene boric acids pinacol ester for detecting peroxynitrite
Colorimetric ratio fluorescent probe in purposes.
Present invention also offers rhodamine compound (the synthesis production of such as 1,6- dihydroxy naphthlenes and 4- lignocaine ketone acids
Thing) preparing the purposes in being used to detect the colorimetric ratio fluorescent probe of peroxynitrite.
The fast super sensitivity high selectivity of the present invention recognizes the notable spy of peroxynitrite colorimetric ratio fluorescent probe
It is that can recognize peroxynitrite to quick high-selectivity to levy, and accurately peroxynitrite quantitatively can be divided
Analysis.
Below will be by the way that the present invention be described in more detail by following examples.Following examples are merely illustrative,
It should be understood that the present invention is not limited by following embodiments.
Embodiment 1
(scheme 1) is by 460mg (1.0mmol) rhodamine compound (such as 1,6- dihydroxy naphthlenes and 4- lignocaine ketone
Acid synthetic product), 297mg (1.0mmol) 4- bromomethyl benzene boric acids pinacol ester, 977mg (3.0mmol) cesium carbonates and
160mg (1.0mmol) KI is dissolved in 6mL dimethylaminos formamide (DMF), then stirring at normal temperature 24h, afterwards extraction revolving
Crude product is obtained, finally using dichloromethane：Methanol (30:1, v/v) system carries out pillar layer separation, obtains pure product
569mg, yield is 87 ﹪.
(scheme 2) is by 460mg (1.0mmol) rhodamine compound (such as 1,6- dihydroxy naphthlenes and 4- lignocaine ketone
Acid synthetic product), 356.4mg (1.2mmol) 4- bromomethyl benzene boric acids pinacol ester, 977mg (3.0mmol) cesium carbonates and
160mg (1.0mmol) KI is dissolved in 6mL dimethylaminos formamide (DMF), then stirring at normal temperature 24h, afterwards extraction revolving
Crude product is obtained, finally using dichloromethane：Methanol (30:1, v/v) system carries out pillar layer separation, obtains pure product
582mg, yield is 89 ﹪.
(scheme 3) is by 460mg (1.0mmol) rhodamine compound (such as 1,6- dihydroxy naphthlenes and 4- lignocaine ketone
Acid synthetic product), 445.5mg (1.5mmol) 4- bromomethyl benzene boric acids pinacol ester, 977mg (3.0mmol) cesium carbonates and
160mg (1.0mmol) KI is dissolved in 6mL dimethylaminos formamide (DMF), then stirring at normal temperature 24h, afterwards extraction revolving
Crude product is obtained, finally using dichloromethane：Methanol (30:1, v/v) system carries out pillar layer separation, obtains pure product
589mg, yield is 90 ﹪.
(scheme 4) is by 460mg (1.0mmol) rhodamine compound (such as 1,6- dihydroxy naphthlenes and 4- lignocaine ketone
Acid synthetic product), 475.2mg (1.6mmol) 4- bromomethyl benzene boric acids pinacol ester, 977mg (3.0mmol) cesium carbonates and
160mg (1.0mmol) KI is dissolved in 6mL dimethylaminos formamide (DMF), then stirring at normal temperature 24h, afterwards extraction revolving
Crude product is obtained, finally using dichloromethane：Methanol (30:1, v/v) system carries out pillar layer separation, obtains pure product
588.6mg, yield is 90 ﹪.
(scheme 5) is by 460mg (1.0mmol) rhodamine compound (such as 1,6- dihydroxy naphthlenes and 4- lignocaine ketone
Acid synthetic product 1), 594mg (2.0mmol) 4- bromomethyl benzene boric acids pinacol ester, 977mg (3.0mmol) cesium carbonates and
160mg (1.0mmol) KI is dissolved in 6mL dimethylaminos formamide (DMF), then stirring at normal temperature 24h, afterwards extraction revolving
Crude product is obtained, finally using dichloromethane：Methanol (30:1, v/v) system carries out pillar layer separation, obtains pure product
582.6mg, yield is 89 ﹪.
Embodiment 2
The present inventor has carried out following test：Various concentrations ONOO-(0-8 μM) is to probe (5 μM) absorption spectrum
Influence and quantitative linearity relation.Said determination is, in 20mM PBS, to be carried out in the pH7.4 aqueous solution containing 10% ethanol
, used probe is prepared probe in embodiment 1, and all spectrum tests are all to add ONOO at 25 DEG C-Make
With what is measured after 3min.As a result referring to Fig. 1.
It will be seen from figure 1 that be gradually increasing with the concentration increase absorption spectrum of peroxynitrite, and solution face
Color is changed into lilac from colourless.
Embodiment 3
Various concentrations ONOO-The influence of (0-2 μM) to probe (5 μM) fluorescence spectrum.Said determination be in 20mM PBS,
Carried out in pH 7.4 aqueous solution containing 10% ethanol, used probe is prepared probe in embodiment 1, and all
Spectrum test is measured after acting on 3min all at 25 DEG C.As a result referring to Fig. 2.
Figure it is seen that along with ONOO in probe solution-Fluorescence spectrum at the increase of concentration, 560nm gradually under
Drop, while the fluorescence spectrum at 630nm gradually rises, and in 0-2 μM of ONOO-In concentration range, ONOO-Concentration and fluorescence
The ratio of intensity shows linear relationship.Therefore, probe of the invention can be accurately determined in sample to be tested using ratio fluorescent method
The content of peroxynitrite.
Embodiment 4
Probe (5 μM) is to ONOO-The test result of (5 μM) response time.Said determination be in 20mM PBS, pH's 7.4
Carried out in the aqueous solution containing 10% ethanol, used probe is prepared probe in embodiment 1, and all spectrum tests
All measured at 25 DEG C.As a result referring to Fig. 3.
From figure 3, it can be seen that add after equivalent peroxynitrite, the very fast rise of fluorescence intensity, and after 5 seconds
Keep maximum.
Embodiment 5
Influence of the different analytes to probe (5 μM) fluorescence spectrum.Wherein：1. blank；2. potassium ion K+；3. calcium ion Ca2 +；4. sodium ion Na+；5. magnesium ion Mg2+；6. aluminium ion Al3+；7. zinc ion Zn2+；8. iron ion Fe3+；9. ferrous ion Fe2+；
10. copper ion Cu2+；11. nitrate ion NO3 -；12. nitrite ion NO2 -；13. cysteine Cys；(3mM) 14. paddy Guang
Sweet peptide GSH (3mM)；15. hydrogen peroxide H2O2(200μM)；16. tertbutanol peroxide TBHP；17. hydroxyl radical free radical OH；18.
Tert-Butanol peroxide free radical OtBu；19. nitric oxide NO；20. singlet oxygen1O2；21. potassium superoxide KO2；22. sodium hypochlorite
NaClO；23. carbonate CO3 2-；24. bisulfite HSO3 -；25. bicarbonate radical HCO3 -；26. bromide ion Br-；27. sulphion
S2-；28. peroxynitrite ONOO-(5 μM), their concentration (in addition to special mark) is 500 μM.All test conditions
It is to carry out in 20mM PBS, pH 7.4 aqueous solution containing 10% ethanol, used probe is prepared in embodiment 1
Probe, and all spectrum tests act on 3min all at 25 DEG C after measure.As a result referring to Fig. 4.Specifically, pipette 50 μ L's
Probe storing solution (1mM) is put into 10mL colorimetric cylinders, then adds 1mL ethanol and 5mL ultra-pure waters, then pipette the 20 above-mentioned analyses of μ L
Thing storing solution is added in colorimetric cylinder, and the PBS solution (pH 7.4,100mM) for then pipetting 2.0mL is added in colorimetric cylinder, is finally used
Ultra-pure water is settled to 10mL.Shake up, stand 3min, you can determine.As a result it is as shown in Figure 4.
, can in specific manner and peroxide from fig. 4, it can be seen that probe has very high selectivity to peroxynitrite
Change nitrite to be reacted, before and after reaction, fluorescence spectrum has a significant change, and other ions acted on probe after fluorescence intensity
Do not occur significant change.
Embodiment six
Probe is shown in Fig. 5, wherein concentration and probe concentration is 20 μM, peroxinitrites to the open hole detection of peroxynitrite
Salinity is respectively 0,10,15,20 μM, test system is ethanol：Water (1:9, v/v), PBS (20mM, pH=7.4) test temperature
For 25 DEG C.
It can be obtained, be continuously increased with the concentration of peroxynitrite by Fig. 5, solution colour is gradually changed into from colourless
Purple, can carry out conveniently open hole detection to peroxynitrite.
Although with above embodiments describing the present invention, it should be appreciated that before the spirit without departing substantially from the present invention
Put, the present invention further can be modified and changed, and these modification and variation belong to protection scope of the present invention it
It is interior.
Claims (10)
1. compound, it has following structure
In above formula：R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12, R13, R14, R15And R16For hydrogen atom, straight or branched
Alkyl, straight or branched alkoxyl, sulfonic group, ester group, carboxyl；R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12, R13,
R14, R15And R16Can be with identical or different.
2. compound according to claim 1, it is the compound of following structure：
3. the preparation for detecting peroxynitrite content in sample, it includes the compound with following structure：
In above formula：R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12, R13, R14, R15And R16For hydrogen atom, straight or branched
Alkyl, straight or branched alkoxyl, sulfonic group, ester group, carboxyl；R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12, R13,
R14, R15And R16Can be with identical or different.
4. preparation according to claim 3, wherein described compound is：
5. preparation according to claim 3, wherein described sample is river sample.
6. the kit for detecting peroxynitrite content in sample, it includes the compound with following structure：
In above formula：R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12, R13, R14, R15And R16For hydrogen atom, straight or branched
Alkyl, straight or branched alkoxyl, sulfonic group, ester group, carboxyl；R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, R12, R13,
R14, R15And R16Can be with identical or different.
7. kit according to claim 6, wherein described compound is：
8. kit according to claim 6, wherein described sample is river sample.
9. the kit according to any one of claim 6-8, it is also including the use of specification.
10. the kit according to any one of claim 6-8, it also includes being used to detect peroxidating nitrous in sample
The buffer of phosphate content.
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| CN115894426A (en) * | 2022-11-14 | 2023-04-04 | 济南大学 | Fluorescent probe for high-selectivity sensitive detection of carbon monoxide, preparation method and application |
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