CN115246827B - Cu (copper) alloy 2+ Sensitive hydrazone derivative and preparation and application thereof - Google Patents
Cu (copper) alloy 2+ Sensitive hydrazone derivative and preparation and application thereof Download PDFInfo
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- CN115246827B CN115246827B CN202111585645.2A CN202111585645A CN115246827B CN 115246827 B CN115246827 B CN 115246827B CN 202111585645 A CN202111585645 A CN 202111585645A CN 115246827 B CN115246827 B CN 115246827B
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- 239000010949 copper Substances 0.000 title claims abstract description 95
- 150000007857 hydrazones Chemical class 0.000 title claims abstract description 59
- 238000002360 preparation method Methods 0.000 title claims description 25
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 title claims description 5
- 229910052802 copper Inorganic materials 0.000 title claims description 5
- 239000000956 alloy Substances 0.000 title claims description 3
- 229910045601 alloy Inorganic materials 0.000 title claims description 3
- 239000007850 fluorescent dye Substances 0.000 claims abstract description 28
- DMZATVKONWVLBE-UHFFFAOYSA-N (4-nitro-2,1,3-benzoxadiazol-7-yl)hydrazine Chemical compound NNC1=CC=C([N+]([O-])=O)C2=NON=C12 DMZATVKONWVLBE-UHFFFAOYSA-N 0.000 claims abstract description 25
- GBWMIOYSMWCYIZ-UHFFFAOYSA-N 6-methyl-4-oxochromene-3-carbaldehyde Chemical compound O1C=C(C=O)C(=O)C2=CC(C)=CC=C21 GBWMIOYSMWCYIZ-UHFFFAOYSA-N 0.000 claims abstract description 8
- 229910021645 metal ion Inorganic materials 0.000 claims description 22
- 239000002904 solvent Substances 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 9
- 239000003960 organic solvent Substances 0.000 claims description 9
- 239000012043 crude product Substances 0.000 claims description 4
- 238000010992 reflux Methods 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims 1
- 239000000523 sample Substances 0.000 abstract description 54
- IGHBXJSNZCFXNK-UHFFFAOYSA-N 4-chloro-7-nitrobenzofurazan Chemical compound [O-][N+](=O)C1=CC=C(Cl)C2=NON=C12 IGHBXJSNZCFXNK-UHFFFAOYSA-N 0.000 abstract description 13
- 238000010521 absorption reaction Methods 0.000 abstract description 12
- 238000001514 detection method Methods 0.000 abstract description 11
- NWZSZGALRFJKBT-KNIFDHDWSA-N (2s)-2,6-diaminohexanoic acid;(2s)-2-hydroxybutanedioic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O.NCCCC[C@H](N)C(O)=O NWZSZGALRFJKBT-KNIFDHDWSA-N 0.000 abstract description 9
- JPVYNHNXODAKFH-UHFFFAOYSA-N Cu2+ Chemical compound [Cu+2] JPVYNHNXODAKFH-UHFFFAOYSA-N 0.000 abstract description 9
- 229910001431 copper ion Inorganic materials 0.000 abstract description 9
- IKDUDTNKRLTJSI-UHFFFAOYSA-N hydrazine monohydrate Substances O.NN IKDUDTNKRLTJSI-UHFFFAOYSA-N 0.000 abstract description 9
- 230000007613 environmental effect Effects 0.000 abstract description 6
- 239000002994 raw material Substances 0.000 abstract description 6
- 239000000126 substance Substances 0.000 abstract description 5
- 239000002184 metal Substances 0.000 abstract description 4
- 229910052751 metal Inorganic materials 0.000 abstract description 4
- 150000002739 metals Chemical class 0.000 abstract description 4
- 238000011160 research Methods 0.000 abstract description 4
- 230000000536 complexating effect Effects 0.000 abstract description 3
- 238000004134 energy conservation Methods 0.000 abstract description 3
- 239000013067 intermediate product Substances 0.000 abstract description 3
- 230000002441 reversible effect Effects 0.000 abstract description 3
- 230000002194 synthesizing effect Effects 0.000 abstract description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 42
- 239000000243 solution Substances 0.000 description 35
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 12
- 150000002500 ions Chemical class 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- 238000002474 experimental method Methods 0.000 description 8
- 239000002244 precipitate Substances 0.000 description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 6
- 150000002576 ketones Chemical class 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 230000004044 response Effects 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 238000005481 NMR spectroscopy Methods 0.000 description 5
- 238000001917 fluorescence detection Methods 0.000 description 5
- 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 4
- 238000002835 absorbance Methods 0.000 description 4
- 238000000862 absorption spectrum Methods 0.000 description 4
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 239000008139 complexing agent Substances 0.000 description 3
- 239000001257 hydrogen Substances 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 230000008859 change Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000002189 fluorescence spectrum Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 239000000376 reactant Substances 0.000 description 2
- 239000011541 reaction mixture Substances 0.000 description 2
- 230000003595 spectral effect Effects 0.000 description 2
- 238000004448 titration Methods 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 206010010957 Copper deficiency Diseases 0.000 description 1
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 1
- 239000007995 HEPES buffer Substances 0.000 description 1
- 239000002262 Schiff base Substances 0.000 description 1
- 150000004753 Schiff bases Chemical class 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000004949 mass spectrometry 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
- 239000003068 molecular probe Substances 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 238000002211 ultraviolet spectrum Methods 0.000 description 1
- 238000002371 ultraviolet--visible spectrum Methods 0.000 description 1
- 238000001429 visible spectrum Methods 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 230000008673 vomiting Effects 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
- C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
- C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
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- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
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- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
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- 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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- 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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- 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
- G01N21/6447—Fluorescence; Phosphorescence by visual observation
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- 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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Abstract
The invention relates to a Cu 2+ The sensitive hydrazone derivative is prepared by taking NBD-Cl as a raw material, synthesizing an intermediate product 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole with hydrazine hydrate, and then reacting with 3-formyl-6-methyl chromone, and can be used as a novel copper ion fluorescent probe L. Cu in a plurality of metals is found through research 2+ The fluorescence probe L has good selectivity, and Cu is added 2+ After that, the ultraviolet-visible absorption peak of the fluorescent probe L is red-shifted, and Cu 2+ The linear relation in a certain concentration range is better, and the working curve can know that the probe L and Cu 2+ Complexing in an amount of 1:1 of the substance and the probe is reversible and recyclable. The probe meets the requirements of energy conservation and environmental protection, and can be Cu 2+ Provides a feasible and effective way for identification and detection.
Description
Technical Field
The invention belongs to the technical field of fluorescent molecular probe preparation, and relates to a Cu 2+ Sensitive hydrazone derivatives, and preparation and application thereof are provided.
Background
Copper element is one of trace elements essential to human body and has important functions of promoting cell metabolism, synthesizing oxygen transporting protein and other vital activities. Copper deficiency in human body can cause anemia, coronary heart disease and the like; excessive copper element causes adverse reactions such as vomiting. Therefore, the method is simple to realize and can detect Cu rapidly 2+ There is a great deal of attention to scientists.
The fluorescent probe detection method has the characteristics of good selectivity and high sensitivity, thus being a research hot spot, but has the problems of poor water solubility, long response time, influence of coexisting ions on detection and the like, so that a novel fluorescent probe is developed to realize Cu in water 2+ The detection has important significance.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a Cu for overcoming the defects in the prior art 2+ Sensitive hydrazone derivatives. The compound has good water solubility, short response time to metal ions, and no influence of coexisting ions on detection, and is a good fluorescent probe.
The invention also provides the Cu 2+ The preparation method of the sensitive hydrazone derivative is simple and has low raw material cost.
The invention further provides the Cu 2+ Application of sensitive hydrazone derivatives.
To this end, the invention provides a Cu 2+ The structural formula of the sensitive hydrazone derivative is shown as (I):
the compound of this structure can be called Cu 2+ The sensitive hydrazone derivative has good water solubility, short response time to metal ions, and no influence of coexisting ions on detection, and can be used as a fluorescent probe for detecting metal ions in aqueous solution or organic solvent.
The invention further provides the Cu 2+ A process for the preparation of a sensitive hydrazone derivative comprising: preparation of Cu 2+ Step of sensitive hydrazone derivatives 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole (NBD-NH) 2 ) Mixing with 3-formyl-6-methyl chromone in solvent, and reflux reacting to obtain Cu 2+ Crude sensitive hydrazone derivatives.
In the above reaction, the 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole (NBD-NH) 2 ) The structural formula of the 3-formyl-6-methyl primary ketone is shown as (II) and the structural formula of the 3-formyl-6-methyl primary ketone is shown as (III). Preparation of Cu in the above step 2+ The chemical reaction formula of the sensitive hydrazone derivative is shown in figure 2.
According to the invention, in the preparation of Cu 2+ In the step of the sensitive hydrazone derivative, the 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole (NBD-NH) 2 ) The molar ratio of the 3-formyl-6-methyl primary ketone to the 3-formyl-6-methyl primary ketone is 1: (1.05-1.08), and is preferably 1:1.075.
In one embodiment of the present invention, cu is prepared as described above 2+ In the step of the sensitive hydrazone derivative, the first solvent is an organic solvent, which includes but is not limited to methanol, ethanol, etc., preferably ethanol.
The amount of the first solvent used in the present invention is not particularly limited as long as the reactant can be well dissolved so that the reaction can be smoothly performed.
According to the invention, in the preparation of Cu 2+ The step of separating and purifying Cu is also included after the step of the sensitive hydrazone derivative 2+ Step of sensitive hydrazone derivatives: the prepared Cu 2+ Separating and purifying the crude product of the sensitive hydrazone derivative to prepare Cu 2+ Pure sensitive hydrazone derivative.
For example, in some specific examples, cu is produced 2+ After the crude product (precipitate) of the sensitive hydrazone derivative is cooled to room temperature, the precipitate is washed with ethanol and dried to obtain Cu 2+ Pure sensitive hydrazone derivative.
In the invention, cu is prepared 2+ In the step of the sensitive hydrazone derivative, the 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole (NBD-NH) 2 ) Can be obtained by commercial method or directly prepared.
In one embodiment of the present invention, in preparing Cu 2+ The step of preparing the sensitive hydrazone derivative also comprises the step of preparing 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole (NBD-NH) 2 ) Comprises the steps of: uniformly mixing 4-chloro-7-nitrobenzofurazan (NBD-Cl) solution and 80% hydrazine hydrate solution, stirring for 1h at room temperature to form a tan precipitate, and separating and purifying the reaction product to obtain 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole (NBD-NH) 2 ) The reaction formula is shown in figure 1.
In the invention, the 4-chloro-7-nitrobenzofurazane (NBD-Cl) solution is formed by dissolving 4-chloro-7-nitrobenzofurazane (NBD-Cl) in a II solvent, wherein the II solvent is an organic solvent and comprises but is not limited to chloroform.
In the present invention, the 80% hydrazine hydrate solution is formed by dissolving 80% hydrazine hydrate in a III solvent, wherein the III solvent is an organic solvent, which includes but is not limited to methanol, ethanol, etc.
The amounts of the II-th solvent and the III-th solvent used in the present invention are not particularly limited as long as the reactants can be well dissolved so that the reaction can be smoothly performed.
In one embodiment of the present invention, cu is prepared by the method of the present invention 2+ Sensitive hydrazone derivatives are as follows:
1. preparation of intermediate 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole (NBD-NH) 2 )
(1) 0.1088g of 4-chloro-7-nitrobenzofurazane (NBD-C1) was dissolved in 30mL of chloroform to prepare a solution a.
(2) 0.77mL of hydrazine hydrate was added to 50mL of methanol to form solution b.
(3) Solution b was added to solution a and stirred at room temperature for 1-2h, forming a tan precipitate and separating to form solid c.
2. Preparation of Cu 2+ Sensitive hydrazone derivatives
(1) To 20mL of ethanol was added solid c (0.1040 g of 0.53 mmol).
(2) 3-formyl-6-methyl primary color ketone (0.1074 g, 0.57 mmol) was added to 10mL of ethanol at room temperature
(3) And (3) dropwise adding the solution obtained in the second step into the solution obtained in the first step, stirring, and then heating and refluxing the stirred reaction mixture for 8-10h to obtain a reddish brown solid.
(4) The precipitate was cooled to room temperature, then washed with ethanol and dried to give 0.1325g of the product in 72.6% yield.
The invention also provides the Cu 2+ Application of sensitive hydrazone derivatives as fluorescent probes, based on application of sensitive hydrazone derivatives in Cu in the invention 2+ The sensitive hydrazone derivative is also called a fluorescent probe L or a hydrazone fluorescent probe L.
As can be seen from the above, the molecular structure of the probe L of the invention is shown in the formula I, the synthetic route is shown in the figures 1 and 2, firstly, 4-chloro-7-nitrobenzofurazan (NBD-C1) is taken as a raw material to generate an intermediate product 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole (NBD-NH) with 80% hydrazine hydrate 2 ) Intermediate NBD-NH 2 And reacting with 3-formyl-6-methyl chromone for 8 hours to synthesize the hydrazone probe L.
According to the inventionCu is combined with 2+ The sensitive hydrazone derivative is used as a fluorescent probe for detecting metal ions.
Due to Cu of the invention 2+ The sensitive hydrazone derivative has good water solubility, short response time to metal ions and no influence of coexisting ions on detection, so that the sensitive hydrazone derivative can be used as a fluorescent probe for detecting metal ions in aqueous solution or organic solvent.
In one embodiment of the present invention, the metal ions comprise Cu 2+ 、Hg 2+ 、Fe 3+ The method comprises the steps of carrying out a first treatment on the surface of the Preferably, the metal ion is Cu 2+ 。
The invention also provides the Cu 2+ The application of sensitive hydrazone derivatives in the biological fluorescent stain.
In one embodiment of the present invention, cu produced according to the method of the present invention 2+ The sensitive hydrazone derivative is used for detecting copper ions in water. Cu is added with 2+ Sensitive hydrazone derivative used as fluorescent probe and Cu-containing 2+ Is added to an aqueous solution containing 10% strength -5 ～10 -6 The mol/L fluorescent probe molecules were reacted in HEPES buffer solution of pH 7.4 for 1-5 minutes, and then the system was tested for fluorescence intensity or ultraviolet absorption.
In another embodiment of the invention, cu is produced according to the method of the invention 2+ The sensitive hydrazone derivative is used for detecting copper ions in an organic solvent. Cu is added with 2+ Sensitive hydrazone derivative used as fluorescent probe and Cu-containing 2+ Is added into the organic solvent with the concentration of 10 -5 ～10 -6 The fluorescence intensity or ultraviolet absorption of the system was then tested in a solution of mol/L fluorescent probe molecules in the same organic solvent.
The term "copper ion" as used herein means "Cu 2+ ”。
The term Cu in the present invention 2+ Sensitive hydrazone derivative refers to the hydrazone derivative meeting Cu 2+ A visual color change or a fluorescent reaction or a shift in the ultraviolet absorption peak occurs.
The term "water" in the present invention refers to one or more of natural environmental water, domestic water and medical water without particular limitation and explanation.
The invention synthesizes an intermediate product 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole by using NBD-Cl as a raw material and hydrazine hydrate, and then reacts with 3-formyl-6-methyl chromone to synthesize a novel copper ion fluorescent probe L. Cu in a plurality of metals is found through research 2+ The fluorescence probe L has good selectivity, and Cu is added 2+ After that, the ultraviolet-visible absorption peak of the fluorescent probe L is red-shifted, and Cu 2+ The linear relation in a certain concentration range is better, and the working curve can know that the probe L and Cu 2+ Complexing in an amount of 1:1 of the substance and the probe is reversible and recyclable. The probe meets the requirements of energy conservation and environmental protection, and can be Cu 2+ Provides a feasible and effective way for identification and detection.
Drawings
The invention is described in further detail below with reference to the accompanying drawings:
FIG. 1 shows the chemical reaction scheme for the preparation of the intermediate 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole.
FIG. 2 shows the preparation of Cu 2+ Chemical reaction formula of sensitive hydrazone derivatives.
FIG. 3 shows Cu obtained in the present invention 2+ Hydrogen spectrum analysis chart of sensitive hydrazone derivative.
FIG. 4 shows Cu obtained in the present invention 2+ Mass spectrometry figures of sensitive hydrazone derivatives.
FIG. 5 is an ultraviolet visible spectrum of metal ions added to an ethanol solution of a probe.
FIG. 6 is a fluorescence spectrum of a metal ion added to an ethanol solution of a probe.
FIG. 7 shows the Cu concentration of probe L after adding different ions to ethanol solution 2+ And detecting a fluorescence intensity histogram.
FIG. 8 shows addition of Cu to probe L in ethanol (2. Mu.L) 2+ (0. Mu.L-13. Mu.L) ultraviolet titration chart.
FIG. 9 shows a probe and Cu 2+ Is of the working graph of (2)。
FIG. 10 shows a probe and Cu 2+ Is a reversibility curve of (2).
Detailed Description
In order that the invention may be more readily understood, a detailed description of the invention will be provided below with reference to the accompanying drawings and examples, which are given by way of illustration only and are not limiting the scope of the invention, as to the specific experimental methods not mentioned in the examples below, which are generally carried out in accordance with conventional experimental methods.
Examples
In order that the invention may be more readily understood, the invention will be further described in detail with reference to the following examples, which are given by way of illustration only and are not limiting in scope of application. The starting materials or components used in the present invention may be prepared by commercial or conventional methods unless specifically indicated.
In the following examples, cu was prepared by ultraviolet-visible spectrophotometry (UV 8000, meprida, shanghai) 2+ And carrying out ultraviolet absorption spectrum analysis on the pure product of the sensitive hydrazone derivative.
Preparation of Cu by using a fluorescence spectrophotometer (F7000 type, hitachi, japan) 2+ And (5) performing fluorescence spectrum analysis on the pure sensitive hydrazone derivative.
Example 1: preparation of Cu 2+ Sensitive hydrazone derivatives
1. Preparation of intermediate 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole (NBD-NH) 2 )
(1) 0.1088g of 4-chloro-7-nitrobenzofurazane (NBD-Cl) was dissolved in 30mL of chloroform to form a solution a.
(2) 0.77mL of hydrazine hydrate was added to 50mL of methanol to form solution b.
(3) Solution b was added to solution a and stirred at room temperature for 1-2h, forming a tan precipitate and separating to form solid c.
2. Preparation of Cu 2+ Sensitive hydrazone derivatives
(1) To 20mL of ethanol was added solid c (0.1040 g of 0.53 mmol).
(2) 3-formyl-6-methyl primary color ketone (0.1074 g, 0.57 mmol) was added to 10mL of ethanol at room temperature
(3) And (3) dropwise adding the solution obtained in the second step into the solution obtained in the first step, stirring, and then heating and refluxing the stirred reaction mixture for 8-10h to obtain a reddish brown solid.
(4) The precipitate was cooled to room temperature, then washed with ethanol and dried to give 0.1325g of the product in 72.6% yield.
After the synthesis reaction, cu was prepared by a nuclear magnetic resonance apparatus (Bruker AVANCE 300MHz type nuclear magnetic resonance apparatus, bruker Co., switzerland) 2+ Nuclear magnetic resonance analysis is carried out on the pure product of the sensitive hydrazone derivative, and the result is as follows:
1H NMR(300MHz，DMSO-D6)δ13.01，9.07，8.73，8.59，7.86，7.66，7.20，2.50，2.45。
the kind of the hydrogen atoms in the organic matters contained in the sample in the chemical environment can be deduced through a nuclear magnetic resonance hydrogen spectrogram (see figure 3), so that the structure of the detected sample can be deduced, and the nuclear magnetic resonance hydrogen spectrogram shown in figure 3 shows that the sample contains the novel Schiff base fluorescent probe which is the target product of the invention, but also contains the raw material 3-formyl-6-methyl chromone, which indicates that the reaction is incomplete due to the problem in the experimental operation.
The molecular weight of the sample is measured by nuclear magnetic resonance spectrum (see figure 4), the theoretical molecular weight of the probe L+Na is 388.3050, the molecular weight of the probe L+Na in the sample is 388.0621, and the target product can be determined to be successfully synthesized by analyzing the invention.
Example 2: investigation of color Change in the addition of Metal ions to the ethanol solution of the Probe
Ion selectivity is one of the very important evaluation indicators in fluorescent probe performance. By mixing various metal ions Al 3+ ，Cd 2+ ，Co 2+ ，Cr 3+ ，Sr 2+ ，Cu 2+ ，Na + ，Ni 2+ ，K + And Zn 2+ Respectively adding the fluorescent probes into ethanol solution of the fluorescent probes L, and performing spectral detection on the sample. As a result, as shown in FIG. 5, it was found that the ethanol solution of probe L (blank) had an absorption peak at 495nm, and absorbance (Abs)Point) is 0.1415, the absorbance peak and absorbance are substantially unchanged when other metal ions are added. But Cu is 2+ After the addition, the color of the solution changed, and the peak of the ultraviolet absorption spectrum at 495nm disappeared, and then the wavelength of the absorption peak was red-shifted to 512nm. The data detected by ultraviolet and visible spectrum can obtain the Cu of the probe L 2+ The ion selectivity and the recognition performance are good.
In the fluorescence detection of the selectivity experiment, probe L shows a reaction to Cu 2+ Is a function of the spectral response of (a). Fluorescence detection excitation wavelength: 490nm, emission wavelength: 500nm, slit: 20nm. At this time, probe L had a peak at 555nm, and Cu alone was added in the selection of many metals 2+ The post peak was 534nm and the shift occurred. And at the time of adding Cu 2+ Fluorescence of the system is reduced after post-fluorescence, cu 2 + The fluorescence of probe L can be quenched, see FIG. 6.
Example 3: competing with each other
The ultraviolet-visible absorption spectrum can reflect the interaction of the probe molecules and the metal ions, and it is very important that the fluorescent probe can singly identify a certain metal ion, and if other ions interfere with the probe, the practicability of the probe can be greatly reduced, so that the invention needs to correspondingly explore the ion competitiveness. In order to explore the practical application capability of the probe, the invention aims at probe L and Cu 2+ Ion competition experiments were performed for the interactions of (c).
Under the condition of ensuring complete consistency, the invention analyzes various metal ion pair probes and Cu 2+ The reaction was affected. As shown in FIG. 7, the present invention compares the sample+metal ion+Cu 2+ The system and the sample+metal ion system are subjected to ultraviolet visible absorption, and under the condition of 583.5nm wavelength, other metal ions exist and only Cu exists in the system 2+ The resulting changes are substantially identical. Experimental data results show that the fluorescent probe L has stronger anti-interference capability in ethanol solution, and the probe pair Cu 2+ Is hardly affected by other metal ions.
Example 4: titration experiments
With concentrationIs 1.0X10 -5 Cu in mol/L 2+ Solution Detitrating Probe L (1.0X10) -5 mol/L), as shown in FIG. 8, when Cu 2+ As the concentration of (2) increases gradually from 0 μm, the ultraviolet-visible absorption spectrum of the solution increases in absorption at 602nm and decreases in absorption at 535 nm. When Cu is 2+ After addition to 11. Mu.L, there was substantially no increase at 602nm and substantially no decrease at 535 nm.
Example 5:
to further determine Cu 2+ The coordination with the probe was analyzed by continuously varying the amount of the same substance, and the result is shown in FIG. 9. As can be seen from FIG. 9, with Cu 2+ The concentration varies in proportion to the concentration of the probe L (1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, 9:1), the absorbance of the probe at the maximum absorption wavelength also varies with their variation, when Cu 2+ The probe response was maximum at a 1:1 concentration ratio to probe L, and experimental data showed that probe and Cu 2+ The complex ratio of (2) is 1:1.
Example 6:
in order to explore whether the probe accords with the environmental protection concept or not, the probe has reversibility and can be recycled, the invention sets a reversibility experiment, considers EDTA and Cu 2+ The two are relatively stable, so EDTA is adopted as a complexing agent to carry out the detection experiment.
Firstly, preparing the ethanol solution of the probe and carrying out fluorescence detection on the ethanol solution, adding a certain amount of copper ions into the ethanol solution, carrying out fluorescence detection on the copper ions after the reaction is completed, adding EDTA solution with the same concentration as the copper ions into a sample dish and carrying out fluorescence detection on the EDTA solution, and through the three steps, the invention draws a data diagram as shown in figure 10, and discovers that the solution has obvious fluorescence peak rise at 557.6nm after adding complexing agent, but does not return to non-Cu 2+ Presumably because of time or concentration reasons. Meanwhile, in the experiment, the solution after the complexing agent is added can be directly observed through naked eyes to turn from colorless to light pink. Therefore, the invention can obtain that the probe can be reused, and accords with environmental protection and economyThe idea is that.
From the above examples, the invention synthesizes intermediate 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole with hydrazine hydrate by using NBD-Cl as raw material, and then reacts with 3-formyl-6-methyl chromone to synthesize a novel copper ion fluorescent probe L. Cu in a plurality of metals is found through research 2+ The fluorescence probe L has good selectivity, and Cu is added 2+ After that, the ultraviolet-visible absorption peak of the fluorescent probe L is red-shifted, and Cu 2+ The linear relation in a certain concentration range is better, and the working curve can know that the probe L and Cu 2+ Complexing in an amount of 1:1 of the substance and the probe is reversible and recyclable. The probe meets the requirements of energy conservation and environmental protection, and can be Cu 2+ Provides a feasible and effective way for identification and detection.
It should be noted that the above-described embodiments are only for explaining the present invention and do not constitute any limitation of the present invention. The invention has been described with reference to exemplary embodiments, but it is understood that the words which have been used are words of description and illustration, rather than words of limitation. Modifications may be made to the invention as defined in the appended claims, and the invention may be modified without departing from the scope and spirit of the invention. Although the invention is described herein with reference to particular means, materials and embodiments, the invention is not intended to be limited to the particulars disclosed herein, as the invention extends to all other means and applications which perform the same function.
Claims (5)
1. Cu (copper) alloy 2+ The molecular structure of the sensitive hydrazone derivative is shown as a formula (I):
2. a Cu according to claim 1 2+ A process for the preparation of a sensitive hydrazone derivative comprising:
preparation of Cu 2+ The step of the sensitive hydrazone derivative is to mix 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole and 3-formyl-6-methyl chromone in the solvent I and then reflux the mixture to prepare Cu 2+ And (3) a crude product of the sensitive hydrazone derivative, wherein the first solvent is an organic solvent.
3. The preparation method according to claim 2, characterized in that: in the preparation of Cu 2+ In the step of the sensitive hydrazone derivative, the molar ratio of the 4-hydrazino-7-nitro-2, 1, 3-benzoxadiazole to the 3-formyl-6-methyl chromone is 1 (1.05-1.08).
4. A method of preparation according to claim 2 or 3, characterized in that:
in the preparation of Cu 2+ The step of separating and purifying Cu is also included after the step of the sensitive hydrazone derivative 2+ Step of sensitive hydrazone derivatives: the prepared Cu 2+ Separating and purifying the crude product of the sensitive hydrazone derivative to prepare Cu 2+ Pure sensitive hydrazone derivative.
5. The Cu of claim 1 2+ Application of sensitive hydrazone derivatives in preparation of fluorescent probes; the application is to make Cu 2+ The sensitive hydrazone derivative is used as a fluorescent probe for detecting metal ions; the metal ion is Cu 2+ 。
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