CN107840855B - fluorescent probe and application thereof - Google Patents
fluorescent probe and application thereof Download PDFInfo
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- CN107840855B CN107840855B CN201711067277.6A CN201711067277A CN107840855B CN 107840855 B CN107840855 B CN 107840855B CN 201711067277 A CN201711067277 A CN 201711067277A CN 107840855 B CN107840855 B CN 107840855B
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- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 25
- 238000001514 detection method Methods 0.000 claims abstract description 17
- 239000000523 sample Substances 0.000 claims description 21
- 239000000243 solution Substances 0.000 claims description 17
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 15
- 239000012086 standard solution Substances 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 3
- CWLKGDAVCFYWJK-UHFFFAOYSA-N 3-aminophenol Chemical compound NC1=CC=CC(O)=C1 CWLKGDAVCFYWJK-UHFFFAOYSA-N 0.000 abstract description 5
- 229940018563 3-aminophenol Drugs 0.000 abstract description 3
- XBDQKXXYIPTUBI-UHFFFAOYSA-N Propionic acid Substances CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 abstract description 3
- 150000001875 compounds Chemical class 0.000 abstract description 2
- 238000006482 condensation reaction Methods 0.000 abstract description 2
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 4
- 239000012670 alkaline solution Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000007795 chemical reaction product Substances 0.000 description 2
- 238000001917 fluorescence detection Methods 0.000 description 2
- 238000002189 fluorescence spectrum Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000002073 fluorescence micrograph Methods 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
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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
- C07F5/00—Compounds containing elements of Groups 3 or 13 of the Periodic Table
- C07F5/02—Boron compounds
- C07F5/022—Boron compounds without C-boron linkages
-
- 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
- G01N21/6428—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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- 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
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1007—Non-condensed systems
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1044—Heterocyclic compounds characterised by ligands containing two nitrogen atoms as heteroatoms
- C09K2211/1055—Heterocyclic compounds characterised by ligands containing two nitrogen atoms as heteroatoms with other heteroatoms
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- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Optics & Photonics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Materials Engineering (AREA)
- 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 invention discloses a fluorescent probe molecule (I) for detecting alkaline pH, which is prepared by a condensation reaction of a compound 3- (4, 4-difluoro-4-boron-3 a, 4 a-aza-cyclopenta-benzene-8-propionic acid and m-aminophenol, wherein the detection range of the fluorescent probe to the pH is 8.5-11.5, and the optimal detection range is 9.5-11.0.
Description
(I) technical field
the invention relates to the field of chemical analysis and biological analysis, in particular to a fluorescent probe for detecting pH value and a synthesis method and application thereof.
(II) background of the invention
In recent years, a large number of fluorescent probes have been designed and synthesized, and provide a very powerful detection tool for life sciences. Because the fluorescence detection has the obvious advantages of simple operation, suitability for living cell detection and the like, the application of the fluorescence detection in biology is rapidly developed. The most common methods for measuring pH include a pH meter, a pH test paper, etc., which have many limitations in application to biological systems, so that a more efficient pH fluorescent probe needs to be designed for pH detection in biological systems.
The fluorescent probes for detecting alkaline pH are very few, and the probes have very important value in researching extreme microorganisms such as microorganisms living in the environment with pH10.5 (Bacillus pseudobacterius OF4 and the like) or microorganisms in alkaline soil; fluorescent probes can become an important component of a molecular device sensitive to pH, so that development of some fluorescent probes for detecting alkaline pH, particularly probes capable of covering the pH range of 9.0-11.0, is required.
Disclosure of the invention
The invention aims to solve the technical problem of the prior art and provide a fluorescent probe for detecting alkaline pH value.
the technical scheme adopted by the invention is as follows:
A fluorescent probe, characterized in that it has the following structural formula (I):
The fluorescent probe is prepared by a condensation reaction between a compound 3- (4, 4-difluoro-4-boron-3 a, 4 a-aza-cyclopenta-phene-8-propionic acid (formula II) and m-aminophenol, and the reaction formula is as follows:
The solvent of the reaction can be dichloromethane, the condensing agent can be dicyclohexylcarbodiimide and 4-dimethylamino pyridine, the reaction is carried out at room temperature, two main reaction products are obtained, wherein the non-fluorescent product component is the probe I, and the non-fluorescent product component can be obtained by a conventional separation and purification method, such as column chromatography separation and purification.
The fluorescent probe can be used for detecting and analyzing pH in a chemical system or a biological system.
The method for detecting pH by using the fluorescent probe comprises the following steps:
Dissolving the fluorescent probe in DMSO to prepare a detection solution with the concentration of 1 mM; preparing standard solution with pH8.0-pH12.0, adding 1 μ L of the detection solution into each standard solution (1000 μ L), mixing, measuring fluorescence intensity FI, and making standard curve with logarithm of fluorescence intensity (logFI) as ordinate and pH value as abscissa; the pH of the sample to be tested can be determined according to a standard curve.
The detection range of the fluorescent probe for pH is 8.5-11.5, and the optimal detection range is 9.5-11.0.
The molecule I has the advantages of mild preparation conditions, simple and convenient steps and low cost, has a plurality of hydrogen bond donor and acceptor atoms in the molecular structure, can meet the requirement of aqueous solution detection, and has good application prospect.
(IV) description of the drawings
FIG. 1: fluorescence emission spectra of probes (1 μ M) in solutions of different pH at room temperature, with fluorescence intensity on the ordinate and emission wavelength on the abscissa.
FIG. 2: fluorescence intensity-pH curves for probes (1. mu.M) at different pH values at room temperature, with fluorescence intensity on the ordinate and pH on the abscissa.
FIG. 3: the standard curve is prepared according to the fluorescence intensity under different pH values, wherein the ordinate in the graph is the logarithm value of the fluorescence intensity, and the abscissa is the pH value.
FIG. 4: fluorescent images taken under fluorescent light after adding the probe (1. mu.M) to the pH8.5 solution (left) and the pH10.5 solution (right) showed that the probe emits green fluorescence visible to the naked eye in alkaline solution.
(V) detailed description of the preferred embodiments
the technical solution of the present invention is further illustrated by the following specific examples, but the scope of the present invention is not limited thereto:
Detailed Description
the preferred embodiments of the present invention will be described in conjunction with the accompanying drawings, and it will be understood that they are described herein for the purpose of illustration and explanation and not limitation.
Example 1: preparing a probe I:
dissolving 3- (4, 4-difluoro-4-boron-3 a, 4 a-aza-cyclopenta-cene-8-propionic acid (formula II) (0.2mmol), dicyclohexylcarbodiimide (0.2mmol), 4-dimethylaminopyridine (0.02mmol) and 3-aminophenol (0.2mmol) in dry dichloromethane (20ml), stirring at room temperature for 1 hour, and rotary evaporating the solvent to dryness, wherein the main reaction products are two, and the non-fluorescent component is probe I, and the final product is 2mg, and the yield is 2.8%. HRMS (ESI): m/z calcd for (C)18H16BF2N3O2+H)+：356.1382，found 356.1381。
Example 2: measuring fluorescence emission spectra of the fluorescent probe in solutions of different pH:
First, solutions of different pH were prepared, and the probe was dissolved in DMSO and added to the above solutions, respectively, so that the final concentration of the probe was 1. mu.M. The fluorescence intensity was measured by a fluorescence spectrophotometer, and as is clear from FIG. 1, the probe solution showed almost no fluorescence at pH 9.0. As the pH increased to 10.0, the fluorescence intensity of the fluorescent probe began to increase gradually. FIG. 2 shows that the probe has the highest detection sensitivity at a pH around 10.36.
Example 3: the fluorescence probe is used for drawing a standard curve to measure the pH value
Dissolving the fluorescent probe in DMSO to prepare a detection solution with the concentration of 1 mM; preparing standard solution with pH of 8.5-12.0, adding 1 μ L of the detection solution into each standard solution (1000 μ L), mixing, measuring fluorescence intensity FI, and making standard curve with logarithm of fluorescence intensity (logFI) as ordinate and pH value as abscissa (figure 3); the pH of the sample to be tested can be determined according to a standard curve.
Example 4: fluorescent probe detection alkaline solution
Dissolving fluorescence in DMSO to prepare a detection solution with the concentration of 1 mM; after the probe was added to the pH8.5 solution (left) and pH10.5 solution (right) (final concentration 1. mu.M), fluorescence images were taken under fluorescent lamp illumination, and the probe emitted green fluorescence visible to the naked eye in an alkaline solution.
Claims (2)
1. A method for detecting pH value by using a fluorescent probe is characterized by comprising the following steps:
Dissolving the fluorescent probe in DMSO to prepare a detection solution with the concentration of 1 mM; preparing standard solutions with the pH value of 8.0-12.0, respectively adding 1 mu L of detection solution into 1000 mu L of each standard solution, uniformly mixing, and then determining fluorescence intensity FI, wherein a standard curve is prepared by taking the logarithm of the fluorescence intensity as the ordinate and the pH value as the abscissa; measuring the pH value of a sample to be measured according to the standard curve;
The fluorescent probe has the following structural formula (I):
The pH value of the sample to be detected is 8.5-11.5.
2. the method for detecting pH with the fluorescent probe according to claim 1, wherein the pH of the sample to be detected is 9.5-11.0.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711067277.6A CN107840855B (en) | 2017-11-02 | 2017-11-02 | fluorescent probe and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711067277.6A CN107840855B (en) | 2017-11-02 | 2017-11-02 | fluorescent probe and application thereof |
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| Publication Number | Publication Date |
|---|---|
| CN107840855A CN107840855A (en) | 2018-03-27 |
| CN107840855B true CN107840855B (en) | 2019-12-13 |
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| CN201711067277.6A Active CN107840855B (en) | 2017-11-02 | 2017-11-02 | fluorescent probe and application thereof |
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Families Citing this family (1)
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| CN111253420B (en) * | 2018-12-03 | 2021-05-04 | 中国科学院化学研究所 | Pyrene compound pH value fluorescent probe and preparation method and application thereof |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101550156A (en) * | 2009-05-14 | 2009-10-07 | 浙江大学 | Fluorescence probe, synthetic method and uses thereof |
| CN102391854A (en) * | 2011-08-03 | 2012-03-28 | 浙江大学 | Fluorescent chemical sensor for detecting [H plus] and preparation method and usage thereof |
| CN103575714A (en) * | 2013-11-04 | 2014-02-12 | 广东工业大学 | Application of coumarone quinoline compound as difunctional pH probe |
| CN104531139A (en) * | 2015-01-06 | 2015-04-22 | 山西大学 | Carbazole type pH fluorescence probe and preparation method and application thereof |
| CN105548174A (en) * | 2015-12-04 | 2016-05-04 | 贵州大学 | Method using light on/off mode probe to measure solution pH |
| CN106083816A (en) * | 2016-07-06 | 2016-11-09 | 山西大学 | A kind of extremely acid carbazoles pH fluorescent probe and its preparation method and application |
| CN106483113A (en) * | 2016-10-26 | 2017-03-08 | 华南师范大学 | 1 pyrene formic acid is used as the application of pH fluorescence probe |
| CN106800531A (en) * | 2017-01-05 | 2017-06-06 | 山西大学 | A kind of fluorescence probe PMPA and preparation method and application |
-
2017
- 2017-11-02 CN CN201711067277.6A patent/CN107840855B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101550156A (en) * | 2009-05-14 | 2009-10-07 | 浙江大学 | Fluorescence probe, synthetic method and uses thereof |
| CN102391854A (en) * | 2011-08-03 | 2012-03-28 | 浙江大学 | Fluorescent chemical sensor for detecting [H plus] and preparation method and usage thereof |
| CN103575714A (en) * | 2013-11-04 | 2014-02-12 | 广东工业大学 | Application of coumarone quinoline compound as difunctional pH probe |
| CN104531139A (en) * | 2015-01-06 | 2015-04-22 | 山西大学 | Carbazole type pH fluorescence probe and preparation method and application thereof |
| CN105548174A (en) * | 2015-12-04 | 2016-05-04 | 贵州大学 | Method using light on/off mode probe to measure solution pH |
| CN106083816A (en) * | 2016-07-06 | 2016-11-09 | 山西大学 | A kind of extremely acid carbazoles pH fluorescent probe and its preparation method and application |
| CN106483113A (en) * | 2016-10-26 | 2017-03-08 | 华南师范大学 | 1 pyrene formic acid is used as the application of pH fluorescence probe |
| CN106800531A (en) * | 2017-01-05 | 2017-06-06 | 山西大学 | A kind of fluorescence probe PMPA and preparation method and application |
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| Selective molecular imaging of viable cancer cells with pH-activatable fluorescence probes;Yasuteru Urano等;《NATURE MEDICINE》;20081207;第15卷(第1期);第104-109页 * |
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