CN110551498A - Preparation and application of coumarin fluorescent probe for detecting Hg 2+ - Google Patents
Preparation and application of coumarin fluorescent probe for detecting Hg 2+ Download PDFInfo
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- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N coumarin Chemical compound C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 title claims abstract description 11
- 235000001671 coumarin Nutrition 0.000 title claims abstract description 6
- 229960000956 coumarin Drugs 0.000 title claims abstract description 5
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 5
- 238000002360 preparation method Methods 0.000 title abstract description 6
- 150000001875 compounds Chemical class 0.000 claims abstract description 9
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 33
- 239000000243 solution Substances 0.000 claims description 23
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 21
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 14
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 claims description 12
- 239000000047 product Substances 0.000 claims description 12
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 10
- 239000012043 crude product Substances 0.000 claims description 10
- XHXFXVLFKHQFAL-UHFFFAOYSA-N phosphoryl trichloride Chemical compound ClP(Cl)(Cl)=O XHXFXVLFKHQFAL-UHFFFAOYSA-N 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 8
- LTMRRSWNXVJMBA-UHFFFAOYSA-L 2,2-diethylpropanedioate Chemical compound CCC(CC)(C([O-])=O)C([O-])=O LTMRRSWNXVJMBA-UHFFFAOYSA-L 0.000 claims description 6
- 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 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 239000005457 ice water Substances 0.000 claims description 5
- 239000003208 petroleum Substances 0.000 claims description 5
- 239000002904 solvent Substances 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- CFNMUZCFSDMZPQ-GHXNOFRVSA-N 7-[(z)-3-methyl-4-(4-methyl-5-oxo-2h-furan-2-yl)but-2-enoxy]chromen-2-one Chemical compound C=1C=C2C=CC(=O)OC2=CC=1OC/C=C(/C)CC1OC(=O)C(C)=C1 CFNMUZCFSDMZPQ-GHXNOFRVSA-N 0.000 claims description 4
- KZMGYPLQYOPHEL-UHFFFAOYSA-N Boron trifluoride etherate Chemical compound FB(F)F.CCOCC KZMGYPLQYOPHEL-UHFFFAOYSA-N 0.000 claims description 4
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 229960000583 acetic acid Drugs 0.000 claims description 3
- 239000012362 glacial acetic acid Substances 0.000 claims description 3
- 239000012044 organic layer Substances 0.000 claims description 3
- 239000011541 reaction mixture Substances 0.000 claims description 3
- 238000010898 silica gel chromatography Methods 0.000 claims description 3
- 229940101006 anhydrous sodium sulfite Drugs 0.000 claims description 2
- 238000001816 cooling Methods 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 239000003480 eluent Substances 0.000 claims description 2
- 238000001704 evaporation Methods 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 239000011259 mixed solution Substances 0.000 claims description 2
- 238000012544 monitoring process Methods 0.000 claims description 2
- ZJLMKPKYJBQJNH-UHFFFAOYSA-N propane-1,3-dithiol Chemical compound SCCCS ZJLMKPKYJBQJNH-UHFFFAOYSA-N 0.000 claims description 2
- 238000010992 reflux Methods 0.000 claims description 2
- 230000003595 spectral effect Effects 0.000 claims 1
- 238000001514 detection method Methods 0.000 abstract description 13
- 238000004458 analytical method Methods 0.000 abstract description 4
- 230000000694 effects Effects 0.000 abstract description 4
- 238000000034 method Methods 0.000 abstract description 3
- 230000035945 sensitivity Effects 0.000 abstract description 3
- QXAMGWKESXGGNV-UHFFFAOYSA-N 7-(diethylamino)-1-benzopyran-2-one Chemical compound C1=CC(=O)OC2=CC(N(CC)CC)=CC=C21 QXAMGWKESXGGNV-UHFFFAOYSA-N 0.000 abstract description 2
- 238000011065 in-situ storage Methods 0.000 abstract description 2
- 238000005259 measurement Methods 0.000 abstract description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 18
- 239000003068 molecular probe Substances 0.000 description 16
- 230000005284 excitation Effects 0.000 description 6
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 229910052753 mercury Inorganic materials 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 2
- 150000001450 anions Chemical class 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 150000004775 coumarins Chemical class 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000009931 harmful effect Effects 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- 208000028698 Cognitive impairment Diseases 0.000 description 1
- 206010011878 Deafness Diseases 0.000 description 1
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- SMWDFEZZVXVKRB-UHFFFAOYSA-N anhydrous quinoline Natural products N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 1
- 239000012736 aqueous medium Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 208000010877 cognitive disease Diseases 0.000 description 1
- 231100000895 deafness Toxicity 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 238000000295 emission spectrum Methods 0.000 description 1
- 238000002189 fluorescence spectrum Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 208000016354 hearing loss disease Diseases 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 238000006862 quantum yield reaction Methods 0.000 description 1
- -1 quinoline cyclic amine Chemical class 0.000 description 1
- 230000035440 response to pH Effects 0.000 description 1
- 229940001482 sodium sulfite Drugs 0.000 description 1
- 235000010265 sodium sulphite Nutrition 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 229910052723 transition metal Inorganic materials 0.000 description 1
- 150000003624 transition metals Chemical class 0.000 description 1
- 238000012418 validation experiment Methods 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
- C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
- C07D409/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
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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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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1088—Heterocyclic compounds characterised by ligands containing oxygen as the only heteroatom
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- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
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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
- C09K2211/1092—Heterocyclic compounds characterised by ligands containing sulfur 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
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Physics & Mathematics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Optics & Photonics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention relates to a preparation method of a coumarin fluorescent probe and application thereof, wherein the name of the compound is 7- (diethylamino) -3- (1, 3-dithian-2-yl) -2H-pyran-2-one, the compound has small molecular weight, simple structure and very sensitive effect on Hg 2+, shows excellent selectivity and sensitivity, is suitable for naked eye detection of Hg 2+, has the detection limit of 5.0nM, has the potential of in-situ detection as same as the most strict Hg 2+ measurement standard, and provides satisfactory results for the application of the established method in analysis environments and marine product samples.
Description
the invention relates to a preparation method of a coumarin derivative fluorescent molecular probe and application of the coumarin derivative fluorescent molecular probe in Hg 2+ detection.
Background
In recent years, heavy metal and transition metal (HTM) ions have received considerable attention because they have harmful effects on the environment and living systems, among which mercury is a well-known highly toxic substance that has harmful effects on human life and environment even at very low concentrations, it has been reported that Hg 2+ released from natural and industrial sources easily enters biofilms and accumulates in the human body through the food chain, thus seriously impairing human health, such as water overload, deafness and cognitive impairment.
An effective sensitive fluorescent molecular probe is obtained by preparing coumarin derivatives, and a series of experiments show that the fluorescent molecule has good performance and potential application value.
Disclosure of Invention
The invention aims to provide a preparation method and practical application of a novel biological fluorescent probe for naked-eye detection of Hg 2+ by coumarins.
The technical scheme of the invention is as follows:
a quinoline cyclic amine derivative fluorescent molecular probe is characterized by having the following structure:
A method for preparing the coumarin derivative comprises the following steps:
Step 1, adding 4-diethylamino salicylaldehyde into ethanol to be fully dissolved, adding diethyl malonate and piperidine, wherein the mass ratio of the 4-diethylamino salicylaldehyde to the diethyl malonate to the piperidine is 1: 2: 1, refluxing a reaction mixture for 10 hours, monitoring by TLC, removing a solvent under reduced pressure, adding concentrated hydrochloric acid and glacial acetic acid to hydrolyze, stirring at 1() () ° C for 6 hours, cooling the solution to room temperature, pouring into ice water, adding 50% NaOH solution to adjust the pH to 7.0-8.0, adding dichloromethane, washing an organic layer with water for three times, drying with sodium sulfite, concentrating under reduced pressure, purifying an anhydrous crude product through silica gel column chromatography, and obtaining a first-step product, wherein an eluent is the ratio of ethyl acetate to dichloromethane of 1: 8;
Step 2. add DMF to phosphorus oxychloride dropwise at 0 ℃, with a ratio of DMF to phosphorus oxychloride of 6: 5, stirring for 30min to generate a red solution, dissolving the first-step product in DMF, dropwise adding the solution into the red solution, stirring the mixture at 60 ℃ overnight, then pouring the mixture into ice water, adding 50% NaOH solution to adjust the pH until solid is separated out, obtaining a crude product by filtering, washing the crude product with cold ethanol, and then recrystallizing the crude product with ethanol and dichloromethane to obtain a second-step product DAC-CHO;
And 3, dissolving the product obtained in the step 2 and 1, 3-propanedithiol in a dichloromethane solution, adding boron trifluoride etherate, stirring at room temperature for 24 hours, then evaporating in vacuum to remove the solvent, washing the residue with cold petroleum ether for three times, and recrystallizing the obtained solid in a mixed solution of ethyl acetate and petroleum ether at the volume ratio of ethyl acetate to petroleum ether of 1: 3 to obtain the target compound DAC-Hg.
the compound has the advantages that the compound has very sensitive effect on Hg 2+, shows excellent selectivity and sensitivity, is suitable for naked eye detection of Hg 2+ and practical application in various aqueous media such as water, soil and marine products, further shows the potential of measuring and managing HTM ions, and has wide application prospect.
Detailed Description
the present invention is further illustrated in detail by the following examples, but it should be noted that the scope of the present invention is not limited by these examples at all.
the first embodiment is as follows: preparation of 7- (diethylamino) -3- (1, 3-dithian-2-yl) -2H-pyran-2-one
1 6 13 6 17 22 2 2 +5mmol, 965mg 4-diethylaminosalicylaldehyde is added to 20.0mL ethanol and dissolved sufficiently, 10mmol, 1.56mL diethyl malonate and 5mmol, 494 μ L piperidine are added, the amount ratio of the substances 4-diethylaminosalicylaldehyde to diethyl malonate to piperidine is 1: 2: 1, the reaction mixture is refluxed for 10H, monitored by TLC, the solvent is removed under reduced pressure, 10.0mL concentrated hydrochloric acid and 10.0mL glacial acetic acid are added for hydrolysis, the solution is stirred at 100 ℃ for 6H, the solution is cooled to room temperature and poured into 50mL ice water, 50% NaOH solution is added to adjust the pH to 7.0-8.0, dichloromethane is added, the organic layer is washed three times with water, dried with anhydrous sodium sulfite, concentrated, the crude product is purified by silica gel column chromatography, the ratio of ethyl acetate to dichloromethane is 1: 8, the first product is obtained, 11.7mmol, 902 μ L DMF is added to 9.75, ESI, 907 μ L, the crude product is precipitated as a solution of ethyl acetate to dichloromethane, the mixture is precipitated as a red chloride, 2H, the crude product is added to a solution of the mixture is filtered, the crude product is added dropwise (21H, 24H, 1H, 24H, 1H, 2H, 24H, 1H, 24H, 1H, 8H, 1H, 2H, 1H 8H 1H, 8H, 5H 1H, 5H, 1H 2H 1H 2H, 1H 2H, 5H, 1H 2H, 1H, 5H, 1H, 5H, 1H, 5H, 2H, 1H, 2H, 5H, 2H, 5H, 2H, 1H, 21H, 1H, 5H, 2H, 5H, 1H, 5H, 1H, 5H, 1H, 5H, 21H, 5H, 1H, 5H, 1H, 5H.
The property and application experiment of the fluorescent molecular compound, the fluorescent molecular probe prepared in the first embodiment is tested in the second to eighth embodiments, and the specific data and analysis are as follows:
Example two:
FIG. 1 mechanism study of Hg 2+ detection by target compound
example three:
FIG. 2 is a graph of UV/Vis absorption spectra of the fluorescent molecular probes and their reaction with Hg 2+ in PBS buffer
Emission spectra of free DAC-Hg (10. mu.M) in PBS buffer (10mM, 10% DMSO, pH7.4) were collected 25 times. The excitation wavelength was 400nm, the photomultiplier voltage was set at 500V, the slit width was set at 5.0nm, and detection was performed on an Shimadzu UV-2550 instrument.
the results show that free probe DAC-Hg showed a peak at 403nm, while both DAC-CHO and DACHg + Hg 2+ showed peaks at 444nm the subsequently determined fluorescence quantum yield of DAC-Hg was 0.93 and the assay product DAC-CHO was 0.27.
Example four:
FIG. 3: fluorescence intensity of DAC-Hg and its response to pH environment
mu.M fluorescent molecular probe was dissolved in PBS (pH 7.4, 10mM, 10% DMSO), and the probe was subjected to different pH values to measure the change in fluorescence intensity, with the pH set in the range of 2-12. The excitation wavelength was 400nm, the photomultiplier voltage was set at 500V, and the slit width was set at 5.0 nm.
Both substances show stability over a wide pH range, which is advantageous for practical use.
Example five:
FIG. 4: in PBS solution, the fluorescence spectrum of the fluorescent molecular probe responding with time
mu.M fluorescent molecular probe was dissolved in PBS (pH 7.4, 10mM, 10% DMSO) and its performance was measured at different times, respectively, set in the range of 0-24 h. The excitation wavelength was 400nm, the photomultiplier voltage was set at 500V, and the slit width was set at 5.0 nm.
the results show that the reaction was completed within 2 hours and remained above 12h, indicating the stability of the fluorescent molecular probe detection system.
Example six:
FIG. 5: FT-IR Spectroscopy validation experiment in PBS solution
the assay was performed by dissolving 10. mu.M fluorescent molecular probe in PBS (pH 7.4, 10mM, 10% DMSO) and incubating at 37 ℃. The excitation wavelength was 400nm, the photomultiplier voltage was set at 500V, and the slit width was set at 5.0 nm.
The results show that the absorption signals at 1722cm-1 and 3125cm-1 correspond to stretching vibrations (influenced by coumarin structure) of-C ═ O, -CH2-CH2-CH 2-in DAC-Hg, respectively, the addition of Hg 2+ causes the appearance of typical carbonyl absorption peaks at 2972cm-1, 1683cm-1, 1635cm-1, 1581cm-1, 1523cm-1, and the disappearance of-C ═ C-C ═ O at 3125cm-1, the graph of DAC-Hg + Hg 2+ is almost identical to the spectrum DAC-CHO, indicating that deprotection reactions of DAC-Hg with Hg 2+ were successful.
Example seven:
FIG. 6 shows the selectivity of the fluorescent molecular probe for Hg 2+ in PBS solution among different amino acids and anions
mu.M fluorescent molecular probe was dissolved in PBS (pH 7.4, 10mM, 10% DMSO) and its performance was measured under different amino acids and anions, respectively, at a concentration of 1.0 mM. The excitation wavelength was 400nm, the photomultiplier voltage was set at 500V, and the slit width was set at 5.0 nm.
2+All these results indicate that DAC-Hg is able to show a "turn-off" effect with high selectivity without significant interference from other metal ions or amino acids.
Example eight:
FIG. 7 Effect of different sequences of EDTA addition on the irreversible mechanism of Hg 2+ in DAC-Hg
the fluorescence intensity of the fluorescent molecular probe was measured by dissolving 10. mu.M of the fluorescent molecular probe in a PBS (pH 7.4, 10mM, 10% DMSO) solution and changing the order of addition of EDTA. The excitation wavelength was 400nm, the photomultiplier voltage was set at 500V, and the slit width was set at 5.0 nm.
It can be seen from the figure that the addition of high concentrations of EDTA (200. mu.M) after complete incubation of the detection system including DAC-Hg and Hg 2+ did not interfere with the detection results.
Example nine:
FIG. 8: analysis of Hydrogen and carbon spectra of the Compound of interest of the present invention
The results show that in practical application, the fluorescent molecular probe provided by the invention has high selectivity and sensitivity, can be used for detecting Hg 2+ with the naked eye, has the detection limit of 5.0nM which is higher than that of other metal ions when Hg 2+ is turned off, has the potential of in-situ detection as the strictest Hg 2+ measurement standard, and provides satisfactory results when the established method is applied to analysis environments and marine product samples.
Claims (3)
1. The coumarin fluorescent probe for detecting Hg 2+ is characterized by comprising the following structural formula:
2. The coumarin derivative according to claim 1, prepared by the steps of:
Step 1, adding 4-diethylamino salicylaldehyde into ethanol to be fully dissolved, adding diethyl malonate and piperidine, wherein the mass ratio of the 4-diethylamino salicylaldehyde to the diethyl malonate to the piperidine is 1: 2: 1, refluxing a reaction mixture for 10 hours, monitoring by TLC, removing a solvent under reduced pressure, adding concentrated hydrochloric acid and glacial acetic acid to hydrolyze, stirring at 100 ℃ for 6 hours, cooling the solution to room temperature, pouring into ice water, adding a 50% NaOH solution to adjust the pH value to 7.0-8.0, adding dichloromethane, washing an organic layer with water for three times, drying with anhydrous sodium sulfite, concentrating under reduced pressure, purifying a crude product by silica gel column chromatography, and obtaining a first-step product, wherein an eluent is the ratio of ethyl acetate to dichloromethane of 1: 8;
Step 2, dropwise adding DMF to phosphorus oxychloride at 0 ℃, wherein the ratio of DMF to phosphorus oxychloride is 6: 5, stirring for 30min to generate a red solution, dissolving the product of the first step in DMF, dropwise adding the solution to the red solution, stirring the mixture at 60 ℃ overnight, then pouring the mixture into ice water, adding a 50% NaOH solution to adjust the pH value until solid is precipitated, filtering to obtain a crude product, washing with cold ethanol, and then recrystallizing with ethanol and dichloromethane to obtain a product of the second step;
And 3, dissolving the product obtained in the step 2 and 1, 3-propanedithiol in a dichloromethane solution, adding boron trifluoride etherate, stirring at room temperature for 24 hours, then evaporating in vacuum to remove the solvent, washing the residue with cold petroleum ether for three times, and recrystallizing the obtained solid in a mixed solution of ethyl acetate and petroleum ether at the volume ratio of 1: 3 to obtain the target compound.
3. The spectral characteristics and practical applications of the fluorescent molecule according to claims 1 and 2.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810577524.5A CN110551498A (en) | 2018-06-04 | 2018-06-04 | Preparation and application of coumarin fluorescent probe for detecting Hg 2+ |
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| CN201810577524.5A CN110551498A (en) | 2018-06-04 | 2018-06-04 | Preparation and application of coumarin fluorescent probe for detecting Hg 2+ |
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| CN201810577524.5A Pending CN110551498A (en) | 2018-06-04 | 2018-06-04 | Preparation and application of coumarin fluorescent probe for detecting Hg 2+ |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112391161A (en) * | 2020-11-30 | 2021-02-23 | 齐鲁工业大学 | Fluorescent probe based on coumarin derivative and preparation method thereof |
| CN113004256A (en) * | 2021-02-26 | 2021-06-22 | 三峡大学 | Ratio type probe for detecting mercury ions and preparation method and application thereof |
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| KR20110020454A (en) * | 2009-08-24 | 2011-03-03 | 고려대학교 산학협력단 | Coumarin derivatives having hg(ii) ion selectivity, method for preparing therefor, detecting method using the same |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| KR20110020454A (en) * | 2009-08-24 | 2011-03-03 | 고려대학교 산학협력단 | Coumarin derivatives having hg(ii) ion selectivity, method for preparing therefor, detecting method using the same |
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| CHAO SONG ET AL.,: "Fluorescent theranostic agents for Hg2+ detection and detoxification treatment", 《CHEMCOMM》 * |
| JA HYUNG KIM ET AL.，: "Fluorescent coumarinyldithiane as a selective chemodosimeter for mercury(II) ion in aqueous solution", 《TETRAHEDRON LETTERS》 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112391161A (en) * | 2020-11-30 | 2021-02-23 | 齐鲁工业大学 | Fluorescent probe based on coumarin derivative and preparation method thereof |
| CN113004256A (en) * | 2021-02-26 | 2021-06-22 | 三峡大学 | Ratio type probe for detecting mercury ions and preparation method and application thereof |
| CN113004256B (en) * | 2021-02-26 | 2022-08-05 | 三峡大学 | Ratio type probe for detecting mercury ions and preparation method and application thereof |
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