CN110373181A - A kind of fluorescence probe detecting fluorine ion - Google Patents
A kind of fluorescence probe detecting fluorine ion Download PDFInfo
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- CN110373181A CN110373181A CN201910613665.2A CN201910613665A CN110373181A CN 110373181 A CN110373181 A CN 110373181A CN 201910613665 A CN201910613665 A CN 201910613665A CN 110373181 A CN110373181 A CN 110373181A
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- fluorine ion
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- fluorescence probe
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- KRHYYFGTRYWZRS-UHFFFAOYSA-M Fluoride anion Chemical compound [F-] KRHYYFGTRYWZRS-UHFFFAOYSA-M 0.000 title claims abstract description 48
- 239000000523 sample Substances 0.000 title claims abstract description 40
- 239000000126 substance Substances 0.000 claims abstract description 5
- 238000001514 detection method Methods 0.000 claims description 12
- 238000006243 chemical reaction Methods 0.000 abstract description 28
- 230000015572 biosynthetic process Effects 0.000 abstract description 10
- 238000003786 synthesis reaction Methods 0.000 abstract description 10
- PQMOXTJVIYEOQL-UHFFFAOYSA-N Cumarin Natural products CC(C)=CCC1=C(O)C(C(=O)C(C)CC)=C(O)C2=C1OC(=O)C=C2CCC PQMOXTJVIYEOQL-UHFFFAOYSA-N 0.000 abstract description 4
- FSOGIJPGPZWNGO-UHFFFAOYSA-N Meomammein Natural products CCC(C)C(=O)C1=C(O)C(CC=C(C)C)=C(O)C2=C1OC(=O)C=C2CCC FSOGIJPGPZWNGO-UHFFFAOYSA-N 0.000 abstract description 4
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N coumarin Chemical compound C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 abstract description 4
- MHYGQXWCZAYSLJ-UHFFFAOYSA-N tert-butyl-chloro-diphenylsilane Chemical compound C=1C=CC=CC=1[Si](Cl)(C(C)(C)C)C1=CC=CC=C1 MHYGQXWCZAYSLJ-UHFFFAOYSA-N 0.000 abstract description 3
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 abstract 2
- 125000006289 hydroxybenzyl group Chemical group 0.000 abstract 1
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 27
- 210000004027 cell Anatomy 0.000 description 16
- 150000001875 compounds Chemical class 0.000 description 10
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 229910052731 fluorine Inorganic materials 0.000 description 9
- 239000011737 fluorine Substances 0.000 description 9
- 239000007788 liquid Substances 0.000 description 9
- 239000012043 crude product Substances 0.000 description 8
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 7
- 150000002500 ions Chemical class 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- YCKRFDGAMUMZLT-UHFFFAOYSA-N Fluorine atom Chemical compound [F] YCKRFDGAMUMZLT-UHFFFAOYSA-N 0.000 description 6
- 239000005457 ice water Substances 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 238000000034 method Methods 0.000 description 5
- RGHHSNMVTDWUBI-UHFFFAOYSA-N 4-hydroxybenzaldehyde Chemical compound OC1=CC=C(C=O)C=C1 RGHHSNMVTDWUBI-UHFFFAOYSA-N 0.000 description 4
- MUDSDYNRBDKLGK-UHFFFAOYSA-N 4-methylquinoline Chemical compound C1=CC=C2C(C)=CC=NC2=C1 MUDSDYNRBDKLGK-UHFFFAOYSA-N 0.000 description 4
- 235000004237 Crocus Nutrition 0.000 description 4
- 241000596148 Crocus Species 0.000 description 4
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 4
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 4
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 4
- 238000000605 extraction Methods 0.000 description 4
- -1 fluorine ions Chemical class 0.000 description 4
- PUZPDOWCWNUUKD-UHFFFAOYSA-M sodium fluoride Chemical compound [F-].[Na+] PUZPDOWCWNUUKD-UHFFFAOYSA-M 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- HJUGFYREWKUQJT-UHFFFAOYSA-N tetrabromomethane Chemical compound BrC(Br)(Br)Br HJUGFYREWKUQJT-UHFFFAOYSA-N 0.000 description 4
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 238000003384 imaging method Methods 0.000 description 3
- 239000003208 petroleum Substances 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 239000000741 silica gel Substances 0.000 description 3
- 229910002027 silica gel Inorganic materials 0.000 description 3
- 238000010898 silica gel chromatography Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 238000010792 warming Methods 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 2
- 150000001450 anions Chemical class 0.000 description 2
- 210000000988 bone and bone Anatomy 0.000 description 2
- 208000004042 dental fluorosis Diseases 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 239000003068 molecular probe Substances 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 239000012074 organic phase Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000011775 sodium fluoride Substances 0.000 description 2
- 235000013024 sodium fluoride Nutrition 0.000 description 2
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 2
- RIOQSEWOXXDEQQ-UHFFFAOYSA-N triphenylphosphine Chemical compound C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 RIOQSEWOXXDEQQ-UHFFFAOYSA-N 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- KRZIKXRXQZBATP-UHFFFAOYSA-N 2,3,4,5-tetramethylquinoline Chemical compound C1=CC(C)=C2C(C)=C(C)C(C)=NC2=C1 KRZIKXRXQZBATP-UHFFFAOYSA-N 0.000 description 1
- 206010016818 Fluorosis Diseases 0.000 description 1
- 239000002879 Lewis base Substances 0.000 description 1
- 206010029164 Nephrotic syndrome Diseases 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 235000010290 biphenyl Nutrition 0.000 description 1
- 239000004305 biphenyl Substances 0.000 description 1
- 125000006267 biphenyl group Chemical group 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 238000012875 competitive assay Methods 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 208000002925 dental caries Diseases 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000005447 environmental material Substances 0.000 description 1
- 150000002170 ethers Chemical class 0.000 description 1
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 150000007527 lewis bases Chemical class 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 208000009928 nephrosis Diseases 0.000 description 1
- 231100001027 nephrosis Toxicity 0.000 description 1
- ZUOUZKKEUPVFJK-UHFFFAOYSA-N phenylbenzene Natural products C1=CC=CC=C1C1=CC=CC=C1 ZUOUZKKEUPVFJK-UHFFFAOYSA-N 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 1
Classifications
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- 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
- C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic Table
- C07F7/02—Silicon compounds
- C07F7/08—Compounds having one or more C—Si linkages
- C07F7/18—Compounds having one or more C—Si linkages as well as one or more C—O—Si linkages
- C07F7/1804—Compounds having Si-O-C linkages
-
- 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
- C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic Table
- C07F7/02—Silicon compounds
- C07F7/08—Compounds having one or more C—Si linkages
- C07F7/18—Compounds having one or more C—Si linkages as well as one or more C—O—Si linkages
- C07F7/1804—Compounds having Si-O-C linkages
- C07F7/1872—Preparation; Treatments not provided for in C07F7/20
- C07F7/188—Preparation; Treatments not provided for in C07F7/20 by reactions involving the formation of Si-O linkages
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- 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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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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
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
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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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- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The invention discloses a kind of fluorescence probe Mito-PF for detecting fluorine ion, belong to technical field of analytical chemistry.The fluorescence probe is connector to hydroxybenzyl using a kind of hemicyanine dye containing cumarin and quinoline as parent, and tert-butyl diphenyl chlorosilane is reaction site, chemical structural formula such as formula () shown in.Fluorescence probe synthesis of the invention is simple, easy to use, can specificity react with fluorine ion and discharge fluorescence, fluorine ion is had good selectivity and anti-interference, the fluorine ion in living cells can be detected.
Description
Technical field
The invention belongs to technical field of analytical chemistry, are related to a kind of fluorescence probe for detecting fluorine ion.
Background technique
Fluorine ion is a kind of substance necessary to human body, is volume minimum, the strongest anion of electronegativity, is one strong
Lewis base, have many unique properties, life science, medicine, chemistry and in terms of have important work
With.Fluorine plays important role in the vital movement of people, and suitable fluorine can play firm bone and tooth, prevent tooth
The surface firm effect again of dissolution is dissolved and is promoted on surface, can with effectively preventing dental caries, treat the osteoporosis of people
Disease etc..Fluorine ion is extremely slow in the intracorporal metabolism of people, and excessive fluorine will cause permanent, irreversible injury to human body, for a long time
The fluorine of excess intake will will lead to the diseases such as den tal fluorosis, fluorosis of bone, nephrosis and uriasis, and furthermore excessive fluorine ion also results in
Blood pressure decline, influences growth and development etc..As people's health awareness has improved, detecting and controlling for fluoride also receives people
Extensive concern and attention.
Currently, people have designed and developed the detection method of a variety of fluorine ions, such as ion selective electrode method, ion chromatography
Method, fluorine reagent colorimetric method and fluorescent molecular probe method etc..Wherein, fluorine ion fluorescent molecular probe is due to highly selective, high
Sensitivity, convenient and efficient, cost is relatively low, the advantages that being detected with real-time in-situ, is subject to the people's attention and widely pays close attention to.
2012, Zhu Weihong seminar report synthesized an example using BODIPY derivative as fluorescent parent, with silicon oxygen bond
As the near infrared fluorescent probe of recognition site, realize to F-Naked eye colorimetric identification and ratio fluorescent detection.
2014, Peng Xiaojun seminar report, which has synthesized an example, can light F in cell mitochondrial-Fluorescence probe.The probe
The F in living cells mitochondria has been lighted for the first time-, can be used to identify intracellular F-Without the interference by other anion.
In conclusion acting in environmental and biological materials to further disclose and probe into fluorine ion, synthesis of selective is designed
By force, the fluorine ion fluorescence probe of high sensitivity has very important theoretical and realistic meaning.
Summary of the invention
The present invention provides a kind of fluorescence probe for being able to detect fluorine ion, and should for the deficiency in the prior art
Probe application detects the imaging applications of fluorine ion in living cells.
A kind of fluorescence probe Mito-PF detecting fluorine ion of the present invention, which is characterized in that the fluorescence probe
Chemical structural formula () shown in:
().
The fluorescence probe Mito-PF of above-mentioned detection fluorine ion is prepared in the following manner:
The synthesis of intermediate A -1: cumarin, 4- methylquinoline, p-methyl benzenesulfonic acid and DMF are added in reaction flask, is warming up to
130 oReaction solution is cooled to room temperature, pours into ice water, be adjusted to pH=7 after reaction 9 hours by C.Methylene chloride extraction, does
Dry, filtering is spin-dried for, obtains crocus oily liquids.Crude product is purified with silica gel column chromatography, obtains crocus powder.
The synthesis of intermediate A -2: parahydroxyben-zaldehyde, tert-butyl diphenyl chlorosilane and three second are added in reaction flask
Amine, methylene chloride make solvent, at room temperature reaction overnight.It extracts after the reaction was completed, dry, concentration obtains gray solid, crude product
It is recrystallized with petroleum ether-ethyl acetate system, compound as white solid 2-1 is obtained by filtration.
Compound 2-1 is added in reaction flask, tetrahydrofuran makees solvent, and under the conditions of ice-water bath, NaBH is added portionwise4,
Reaction overnight at room temperature.Methanol is removed under reduced pressure, washes, methylene chloride extraction dries, filters, is spin-dried for, obtains pale yellow oily liquid
Body.Crude product is purified with silica gel column chromatography, obtains colourless oil liquid compound 2-2.
In reaction flask, carbon tetrabromide is added, anhydrous ether makees solvent, add triphenyl phosphorus, under conditions of ice-water bath,
The diethyl ether solution of compound 2-2 is added, reaction 10 hours is stirred at room temperature.To after completion of the reaction, be removed by filtration solid, rotation
It is dry, intermediate A -2 is obtained, can be directly used in reaction in next step.
The synthesis of probe Mito-PF: in reaction flask, addition intermediate A -1, A-2 and acetonitrile, 90oReaction 20 is small under C
Shi Hou.It is concentrated to get purple oily liquids.Crude product is purified with silica gel column chromatography, obtains probe Mito-PF.
Synthesis of the invention is as follows:
。
The application of fluorescence probe of the present invention fluorine ion in detection living cells.
Fluorescence probe of the present invention can should in detection living cells in fluorine ion, specific detection method are as follows: in Hela
5 μM of Mito-PF is added in cell, is cultivated 15 minutes under 37 oC, cell is substantially without fluorescence.Later by Hela cell and 5
μM Mito-PF cultivated 15 minutes at 37 oC after, with PBS buffer solution wash three times, replace culture medium, respectively by difference
(200 μM, 400 μM) of fluorine ion of concentration are cultivated 15 minutes, and cell issues strong fluorescence.Experiments have shown that Mito-PF can be with
Fluorine ion in effective detection organism, in terms of there is important potential using value.
Beneficial effects of the present invention:
A kind of fluorescence probe detecting fluorine ion according to the present invention, using cumarin and tetramethyl quinoline as parent, with tertiary fourth
Base diphenyl is switch, can be with fluorine ion specific reaction, and fluorescence probe synthesis of the invention is simple, easy to use, can be with
Specificity reacts with fluorine ion discharges fluorescence, not will receive the interference of other ions during detecting fluorine ion, to fluorine
Ion has good selectivity, and can detect to the fluorine ion in living cells, imaging effect is good, and low to cytotoxicity.
Detailed description of the invention
Fig. 1 is the molecular structure of fluorescence probe Mito-PF.
Fig. 2 is fluorescence probe Mito-PF1H NMR spectra.
Fig. 3 is fluorescence probe Mito-PF13C NMR spectra.
Fig. 4 is the uv absorption spectra of fluorescence probe Mito-PF.
Fig. 5 is the selective light spectrogram of fluorescence probe Mito-PF.
Fig. 6 is fluorescence probe Mito-PF with concentration of hydrogen peroxide variation response light spectrogram.
Fig. 7 is fluorescence probe Mito-PF response time spectrogram.
Fig. 8 is fluorescence probe Mito-PF anti-interference test chart.
Fig. 9 is fluorescence probe Mito-PF to the intracellular fluorine ion detection figure of Hela；A. (10 μM) of Mito-PF cultures
Fluorescent image after Hela cell 15 minutes, B. light field, C. coverage diagram, D. are thin by (10 μM) culture Hela of Mito-PF
Fluorescent image after being cultivated 15 minutes by (200 μM) of sodium fluoride again after born of the same parents 15 minutes, E. light field, F. coverage diagram.
Specific embodiment
Present invention will be further explained below with reference to the attached drawings and examples.
Embodiment 1:
The synthesis of intermediate A -1: be separately added into reaction flask cumarin (0.5 g, 2 mmol), 4- methylquinoline (0.35 g,
2.4 mmol), p-methyl benzenesulfonic acid (0.95 g, 5 mmol) and DMF(8 mL).It is warming up to 130oC reacts 9 hours, reaction
After the completion, it is cooled to room temperature, pours into 30 mL ice water, be adjusted to pH=7 with 40% sodium hydroxide solution.Methylene chloride (3 × 30
ML it) extracts, merges organic phase, anhydrous magnesium sulfate is dried, filtered, is spin-dried for, and obtains crocus oily liquids.By crude product silica gel color
Column purification (methylene chloride: methanol=20:1) is composed, obtains crocus powder compounds 3(0.3 g, 40.5%).
The synthesis of intermediate A -2: parahydroxyben-zaldehyde (1 g, 8.2 mmol), TBDPS-Cl are separately added into reaction flask
(2.5 g, 9.1 mmol), triethylamine (1.65 g, 16.3 mmol) and methylene chloride (20 mL), at room temperature reaction overnight.Instead
It after the completion of answering, pours into 1M HCL aqueous solution, methylene chloride (3 × 30 mL) extraction, anhydrous magnesium sulfate is dried, filtered, is spin-dried for
Pale solid, gained crude product recrystallize in 40 mL petroleum ethers, obtain compound as white solid 2-1(2.4 g, and 81%).
Compound 2-1(1.4 g, 3.8 mmol are added in reaction flask) and THF(20 mL), under the conditions of ice-water bath, in batches
NaBH is added4(0.14 g, 3.7 mmol), are gradually heated to room temperature, reaction overnight.Reaction is completed, with methylene chloride (30 mL
× 3) extraction three times, merges organic phase, and anhydrous magnesium sulfate is dry, and vacuum concentration obtains pale yellowish oil liquid.Utilize silica gel chromatograph
Column method purifies (ethyl acetate: petroleum ether=1:4) to resulting crude product, obtains colorless and transparent oily liquids compound (1
G, 71.9%).
Be added in reaction flask carbon tetrabromide (1 g, 3 mmol), anhydrous ether (19 mL) and triphenyl phosphorus (0.63 g,
2.4 mmol), be stirred at room temperature 0.5 h, under ice-water bath, compound 2-2(0.48 g, 1.3 mmol are added into reaction flask), it is extensive
Multiple room temperature, reacts 10 h.After the reaction was completed, it filters, is spin-dried for, obtains oily liquids compound A-2, it is not purified, it is directly used in
In next step.
The synthesis of probe Mito-PF: in reaction flask be added intermediate A -1(0.12 g, 0.35 mmol), intermediate A -2
With acetonitrile (3 mL), it is warming up to 90oC reacts 20 h.After the reaction was completed, it is cooled to room temperature, is concentrated to get purple oily liquids.
By crude product with silica gel chromatographic column purify (methylene chloride: methanol=20: 1), obtaining violet solid Mito-PF(150 mg,
54%).1H NMR (600 MHz, CDCl3) δ 9.80 (s, 1H), 8.65 (d, J = 9.6 Hz, 2H), 8.56
(d, J = 5.2 Hz, 1H), 8.29 (d, J = 2.2 Hz, 1H), 8.06 (d, J = 8.9 Hz, 1H), 8.01
(d, J = 15.4 Hz, 1H), 7.88 (t, J = 7.8 Hz, 1H), 7.77 (t, J = 7.4 Hz, 1H),
7.63 (d, J = 7.0 Hz, 4H), 7.50 (d, J = 8.9 Hz, 1H), 7.39 (t, J = 7.3 Hz, 2H),
7.32 (t, J = 7.4 Hz, 4H), 7.03 (d, J = 8.6 Hz, 2H), 6.70 (d, J = 8.5 Hz, 2H),
6.64 (d, J = 8.8 Hz, 1H), 6.49 (s, 1H), 6.09 (s, 2H), 3.47 (q, J = 7.0 Hz,
4H), 1.26 (t, J = 7.0 Hz, 6H), 1.05 (s, 9H). 13C NMR (151 MHz, CDCl3) δ
160.39, 156.87, 156.25, 154.45, 152.51, 148.10, 147.58, 140.86, 138.13,
135.42, 134.59, 132.34, 131.31, 130.05, 128.73, 128.52, 127.85, 127.06,
126.77, 125.59, 120.60, 119.07, 118.90, 116.14, 114.57, 110.02, 109.48,
96.65, 59.92, 45.20, 26.44, 19.40, 12.58.
The present invention has carried out measure of merit to the probe Mito-PF that embodiment 1 obtains:
1. Mito-PF UV absorption is analyzed
40 equivalent fluorine ion PBS buffer solutions are added in 5 μM of Mito-PF, after being stored at room temperature 30 min, start to test, examine
Result such as Fig. 4 is surveyed, the absorption peak at 500-700 nm reduces, and new absorption peak is generated between 400-500 nm, illustrates probe
Structure changed.
2. Mito-PF is selectively analyzed
40 equivalent fluorine ions are added in 5 μM of Mito-PF or interfering ion is detected after being placed at room temperature for 30 min, examine
Result such as Fig. 5 is surveyed, Mito-PF has strong fluorescence response to fluorine ion, and only has faint response to other ions, it was demonstrated that
Mito-PF has excellent selectivity to fluorine ion.
3. Mito-PF changes response analysis to fluorinion concentration
When the fluorine ion of 0-150 equivalent being added in 5 μM of Mito-PF, fluorescence response intensity is with fluorine ion additional amount
Increase in regularity enhancing, testing result such as Fig. 6, as a result illustrates that Mito-PF is wide to fluorinion concentration detection range and sensitivity
It is high.
4. Mito-PF is to fluorine ion response time analysis
The PBS buffer solution of 40 equivalent fluorine ions is added in 5 μM of Mito-PF, to the response time of Mito-PF and fluorine ion
Detected, testing result such as Fig. 7, the results show that after hydrogen peroxide is added in 0-15 min, Mito-PF to fluorine from
The increase of the fluorescence response intensity of son at any time linearly enhances, and can reach good fluorescence intensity in a relatively short period of time.
The result shows that Mito-PF and fluorine ion response quickly, can be effectively used for the detection of fluorine ion.
5. Mito-PF anti-interference is analyzed
Probe Mito-PF has carried out competitive assay in the presence of the disturbing molecules such as various ions, to detect Mito-PF's
Anti-interference.Testing result such as Fig. 8, in the case where fluorine ion and other ions coexist, Mito-PF still can produce fluorine ion
Raw stable fluorescence response.The above results show that Mito-PF has excellent anti-interference to fluorine ion, can be in other ions
In the presence of efficiently detect fluorine ion.
6. application of the Mito-PF in cell detection
The Mito-PF for being added 10 μM in Hela cell cultivates 15 min, testing result such as Fig. 9, by A in Fig. 9 at 37 oC
Shown, cell is substantially without fluorescence.After Hela cell and 5 μM of Mito-PF are cultivated 15 min at 37 oC by us, use
PBS is washed three times, replaces culture medium, then by (200 μM) 15 min of culture of sodium fluoride buffer solution, as shown in D in Fig. 9, cell
Issue strong fluorescence.Experiment shows that Mito-PF has good imaging to act on the fluorine ion in cell, can be very good to use
In detection organism in fluorine ion, in terms of have important potential using value.
Above-mentioned, although the foregoing specific embodiments of the present invention is described with reference to the accompanying drawings, not to the limit of invention scope
System, the field technical staff should be understood that based on the technical solutions of the present invention those skilled in the art do not need to pay
The various modifications or changes that creative work can be made out are still fallen within the protection scope of the present invention.
Claims (3)
1. it is a kind of detect fluorine ion fluorescence probe, which is characterized in that the fluorescence probe Mito-PF chemical structural formula ()
It is shown:
(I).
2. a kind of fluorescence probe for detecting fluorine ion according to claim 1, which is characterized in that the fluorescence probe
Mito-PF chemical structural formula () shown in.
3. a kind of fluorescence probe for detecting fluorine ion according to claim 1, which is characterized in that the fluorescence probe
Mito-PF can be applied to the detection of fluorine ion in living cells.
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