CN114835642B - Preparation method and application of fluorescent probe for identifying zinc ions and glyphosate in relay manner - Google Patents
Preparation method and application of fluorescent probe for identifying zinc ions and glyphosate in relay manner Download PDFInfo
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- CN114835642B CN114835642B CN202210350350.5A CN202210350350A CN114835642B CN 114835642 B CN114835642 B CN 114835642B CN 202210350350 A CN202210350350 A CN 202210350350A CN 114835642 B CN114835642 B CN 114835642B
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- 239000005562 Glyphosate Substances 0.000 title claims abstract description 110
- 229940097068 glyphosate Drugs 0.000 title claims abstract description 110
- XDDAORKBJWWYJS-UHFFFAOYSA-N glyphosate Chemical compound OC(=O)CNCP(O)(O)=O XDDAORKBJWWYJS-UHFFFAOYSA-N 0.000 title claims abstract description 110
- PTFCDOFLOPIGGS-UHFFFAOYSA-N Zinc dication Chemical compound [Zn+2] PTFCDOFLOPIGGS-UHFFFAOYSA-N 0.000 title claims abstract description 86
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 77
- 238000002360 preparation method Methods 0.000 title abstract description 11
- 238000001514 detection method Methods 0.000 claims abstract description 38
- 239000000523 sample Substances 0.000 claims abstract description 32
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims abstract description 18
- 238000000034 method Methods 0.000 claims abstract description 12
- GCMNJUJAKQGROZ-UHFFFAOYSA-N 1,2-Dihydroquinolin-2-imine Chemical compound C1=CC=CC2=NC(N)=CC=C21 GCMNJUJAKQGROZ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 230000035945 sensitivity Effects 0.000 claims abstract description 5
- NTCCNERMXRIPTR-UHFFFAOYSA-N 2-hydroxy-1-naphthaldehyde Chemical compound C1=CC=CC2=C(C=O)C(O)=CC=C21 NTCCNERMXRIPTR-UHFFFAOYSA-N 0.000 claims abstract description 4
- 238000006482 condensation reaction Methods 0.000 claims abstract description 4
- 239000002689 soil Substances 0.000 claims description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 10
- 230000002441 reversible effect Effects 0.000 claims description 4
- 238000010992 reflux Methods 0.000 claims description 3
- 238000012800 visualization Methods 0.000 claims 1
- 230000008569 process Effects 0.000 abstract description 4
- 238000012360 testing method Methods 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 3
- 230000000007 visual effect Effects 0.000 abstract description 3
- 239000000126 substance Substances 0.000 abstract 1
- 239000011701 zinc Substances 0.000 description 40
- 239000000243 solution Substances 0.000 description 26
- 239000007864 aqueous solution Substances 0.000 description 6
- IAJOBQBIJHVGMQ-UHFFFAOYSA-N 2-amino-4-[hydroxy(methyl)phosphoryl]butanoic acid Chemical compound CP(O)(=O)CCC(N)C(O)=O IAJOBQBIJHVGMQ-UHFFFAOYSA-N 0.000 description 5
- 239000005947 Dimethoate Substances 0.000 description 5
- 239000005561 Glufosinate Substances 0.000 description 5
- 239000005949 Malathion Substances 0.000 description 5
- 238000007405 data analysis Methods 0.000 description 5
- OEBRKCOSUFCWJD-UHFFFAOYSA-N dichlorvos Chemical compound COP(=O)(OC)OC=C(Cl)Cl OEBRKCOSUFCWJD-UHFFFAOYSA-N 0.000 description 5
- 229950001327 dichlorvos Drugs 0.000 description 5
- JXSJBGJIGXNWCI-UHFFFAOYSA-N diethyl 2-[(dimethoxyphosphorothioyl)thio]succinate Chemical compound CCOC(=O)CC(SP(=S)(OC)OC)C(=O)OCC JXSJBGJIGXNWCI-UHFFFAOYSA-N 0.000 description 5
- MCWXGJITAZMZEV-UHFFFAOYSA-N dimethoate Chemical compound CNC(=O)CSP(=S)(OC)OC MCWXGJITAZMZEV-UHFFFAOYSA-N 0.000 description 5
- ZNOLGFHPUIJIMJ-UHFFFAOYSA-N fenitrothion Chemical compound COP(=S)(OC)OC1=CC=C([N+]([O-])=O)C(C)=C1 ZNOLGFHPUIJIMJ-UHFFFAOYSA-N 0.000 description 5
- 229960000453 malathion Drugs 0.000 description 5
- 229960001952 metrifonate Drugs 0.000 description 5
- PZXOQEXFMJCDPG-UHFFFAOYSA-N omethoate Chemical compound CNC(=O)CSP(=O)(OC)OC PZXOQEXFMJCDPG-UHFFFAOYSA-N 0.000 description 5
- LCCNCVORNKJIRZ-UHFFFAOYSA-N parathion Chemical compound CCOP(=S)(OCC)OC1=CC=C([N+]([O-])=O)C=C1 LCCNCVORNKJIRZ-UHFFFAOYSA-N 0.000 description 5
- RLBIQVVOMOPOHC-UHFFFAOYSA-N parathion-methyl Chemical compound COP(=S)(OC)OC1=CC=C([N+]([O-])=O)C=C1 RLBIQVVOMOPOHC-UHFFFAOYSA-N 0.000 description 5
- NFACJZMKEDPNKN-UHFFFAOYSA-N trichlorfon Chemical compound COP(=O)(OC)C(O)C(Cl)(Cl)Cl NFACJZMKEDPNKN-UHFFFAOYSA-N 0.000 description 5
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- YASYVMFAVPKPKE-UHFFFAOYSA-N acephate Chemical compound COP(=O)(SC)NC(C)=O YASYVMFAVPKPKE-UHFFFAOYSA-N 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000000575 pesticide Substances 0.000 description 4
- 230000000536 complexating effect Effects 0.000 description 3
- 230000005284 excitation Effects 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 239000003987 organophosphate pesticide Substances 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000010949 copper Substances 0.000 description 2
- 125000004122 cyclic group Chemical group 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
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- 229910021645 metal ion Inorganic materials 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- -1 thioimine Chemical compound 0.000 description 2
- CNHYKKNIIGEXAY-UHFFFAOYSA-N thiolan-2-imine Chemical compound N=C1CCCS1 CNHYKKNIIGEXAY-UHFFFAOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- KZEVSDGEBAJOTK-UHFFFAOYSA-N 1-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)-2-[5-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]-1,3,4-oxadiazol-2-yl]ethanone Chemical compound N1N=NC=2CN(CCC=21)C(CC=1OC(=NN=1)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)=O KZEVSDGEBAJOTK-UHFFFAOYSA-N 0.000 description 1
- 208000023345 Autoimmune Diseases of the Nervous System Diseases 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 241001541138 Cypho Species 0.000 description 1
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 1
- 208000012902 Nervous system disease Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- FOCVUCIESVLUNU-UHFFFAOYSA-N Thiotepa Chemical compound C1CN1P(N1CC1)(=S)N1CC1 FOCVUCIESVLUNU-UHFFFAOYSA-N 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 208000035362 autoimmune disorder of the nervous system Diseases 0.000 description 1
- DLGYNVMUCSTYDQ-UHFFFAOYSA-N azane;pyridine Chemical compound N.C1=CC=NC=C1 DLGYNVMUCSTYDQ-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 231100000357 carcinogen Toxicity 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 239000013522 chelant Substances 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000001917 fluorescence detection Methods 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 230000002363 herbicidal effect Effects 0.000 description 1
- 239000004009 herbicide Substances 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 238000000967 suction filtration Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 229960001196 thiotepa Drugs 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
- C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
- C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D215/38—Nitrogen atoms
-
- 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"
- 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
-
- 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
- C09K2211/1011—Condensed systems
-
- 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
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the 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
Abstract
A preparation method and application of a fluorescent probe for identifying zinc ions and glyphosate in a relay way relate to the field of analysis and detection. The method solves the problem that the existing fluorescent probe detection method can not identify two substances of zinc ions and glyphosate in a relay way under the same test condition. The preparation method of the fluorescent probe comprises the following steps: and (3) carrying out condensation reaction on the 2-hydroxy-1-naphthaldehyde and the 2-aminoquinoline to obtain the fluorescent probe. The fluorescent probe prepared by the invention can specifically identify zinc ions in DMSO solution, and the complex formed by the probe and the zinc ions can realize selective identification of glyphosate. The identification process has the advantages of good selectivity, strong anti-interference capability, high sensitivity, visual detection, simple operation, low cost and low requirements on equipment and personnel, and provides a new method for relay identification of zinc ions and glyphosate.
Description
Technical Field
The invention relates to the field of analysis and detection, in particular to a preparation method of a fluorescent probe and application of the fluorescent probe in relay recognition of zinc ions and glyphosate.
Background
Zinc is one of trace elements essential to human body, plays an important role in life process, and can be involved in the processes of cell metabolism, apoptosis, gene expression and the like. In human body, both high and low zinc ion content can have an effect on health. When zinc ions are excessively ingested, the ingestion of copper, iron and other elements by the human body is affected. When zinc ion intake is too low, physical retardation, nervous system diseases, immune diseases and the like are caused. In addition, the content of zinc ions also affects the growth and development of plants. Therefore, the method has important significance for detecting zinc ions in the environment and organisms.
Glyphosate is a high-efficiency, non-selective, strong-conductivity and low-cost broad-spectrum herbicide, is widely produced and sold in the global scope, and is the pesticide variety with the largest sales in the global crop protection market. The country is a large country for producing and using glyphosate, so that people with occupational contact are numerous. In recent years, glyphosate has been shown to be potentially toxic to organisms and is re-rated by the World Health Organization (WHO) 2015 as a class 2A carcinogen. The maximum concentration of glyphosate in drinking water is limited to 0.7mg/L and the maximum concentration of glyphosate in fruits is limited to 0.1mg/kg. Therefore, the rapid and convenient detection of the glyphosate in the environment and crops is necessary.
At present, independent identification of zinc ions or glyphosate can be realized, but documents for relay identification of zinc ions and glyphosate on the same detection level are not reported. The invention designs and synthesizes a quinoline fluorescent probe, and is applied to relay identification of zinc ions and glyphosate. The method is simple and convenient to operate, reduces the synthesis work of the probe, improves the detection efficiency, and enables the fluorescent probe detection method to be more energy-saving and green.
Disclosure of Invention
The invention aims to solve the problem that zinc ions and glyphosate are difficult to realize simultaneous detection under the same test condition, and provides a preparation method and application of a fluorescent probe for relay recognition of zinc ions and glyphosate.
The invention discloses a fluorescent probe for relay recognition of zinc ions and glyphosate, which has the molecular structure as follows:
the synthetic route of the fluorescent probe is as follows:
the invention discloses a preparation method of a fluorescent probe for identifying zinc ions and glyphosate in relay, which comprises the following steps: reflux of the 2-hydroxy-1-naphthaldehyde and the 2-aminoquinoline in an ethanol solution is carried out, and condensation reaction is carried out to obtain the fluorescent probe L.
The operation of the fluorescent probe for identifying zinc ions and glyphosate in relay is as follows: in DMSO solution, the probe can detect zinc ion "turn-on", and after glyphosate is directly added into the detection system, the probe and zinc ion complex can detect glyphosate "on-off".
Further, the detection limit of the fluorescent probe on zinc ions is 4.7X10 -7 The detection limit of the probe and zinc ion complex to glyphosate is 2.5X10 mol/L -7 mol/L。
Furthermore, the fluorescent probe can realize visual relay identification with high selectivity, high anti-interference performance and high sensitivity to zinc ions and glyphosate.
Furthermore, the fluorescent probe can realize reversible cycle identification of zinc ions and glyphosate.
When the invention is actually used for detecting the glyphosate-containing sample, the sample is subjected to pretreatment of centrifugation and filtration to remove solid particle impurities.
Furthermore, the complex formed by the fluorescent probe and zinc ions can be applied to the residual detection of glyphosate in soil.
The principle of the invention is as follows:
the fluorescent probe structure prepared by the invention has a larger conjugated system and contains structures such as hydroxyl, imine bond, pyridine nitrogen and the like. These structures can complex with zinc ions, and transfer electrons from the probe to the zinc ions occurs, thus realizing the off-on detection of the zinc ions. The glyphosate molecule contains amino, phosphate, carboxyl and other functional groups, can have strong coordination with zinc ions, and can replace the zinc ions in the probe and zinc ion complex, so that the fluorescence effect is recovered, and the on-off fluorescence detection of the glyphosate is realized. The "off-on-off" principle of the fluorescent probe for relay recognition of zinc ions and glyphosate is shown in figure 1.
Compared with the prior art, the invention has the following beneficial effects:
1) The invention adopts a fluorescent probe detection method for the first time, and realizes relay identification of zinc ions and glyphosate under the same test condition. The invention has good detection effect, simple and convenient operation, reduces the synthesis work of the probe and improves the detection efficiency.
2) The fluorescent probe prepared by the invention has good sensitivity, and the detection limit of zinc ions is 4.7X10 - 7 The detection limit of the probe and zinc ion complex to glyphosate is 2.5X10 mol/L -7 mol/L。
3) The invention applies the fluorescent probe detection method to the residual detection of glyphosate in soil for the first time.
Drawings
FIG. 1 is a schematic diagram of the present invention fluorescent probes for relay recognition of zinc ions and glyphosate;
FIG. 2 preparation of fluorescent probes according to the invention 1 H NMR spectrum;
FIG. 3 preparation of fluorescent probes according to the invention 13 C NMR spectrum;
FIG. 4 is an IR spectrum of a prepared fluorescent probe according to the present invention;
FIG. 5 selectivity for fluorescent probe recognition of metal ions;
FIG. 6 is a graph of the fluorescence response of a fluorescent probe to zinc ions of different concentrations;
FIG. 7 Job's plot curve of fluorescent probe against zinc ions;
FIG. 8 fluorescent probe-Zn 2+ Selectively identifying a relationship diagram for the organophosphorus pesticide;
FIG. 9 fluorescent probe L (1 #) and fluorescent probe L-Zn under ultraviolet light 365nm 2+ (2#)Fluorescent probe L-Zn 2+ Fluorescence change patterns after the action with glyphosate (3 #), trichlorfon (4 #), iminothiolate (5 #), dichlorvos (6 #), malathion (7 #), omethoate (8 #), dimethoate (9 #), acephate (10 #), fenitrothion (11 #), methyl parathion (12 #), parathion (13 #), and glufosinate (14 #), respectively;
FIG. 10 Co-existing organophosphorus pesticide pair fluorescence probe-Zn 2+ Identifying a glyphosate relationship map;
FIG. 11 fluorescent probe-Zn 2+ A graph of fluorescence response to different concentrations of glyphosate;
FIG. 12 fluorescent probe-Zn 2+ Job's plot against glyphosate;
FIG. 13 fluorescence probe pair Zn 2+ And cyclic identification of glyphosate;
Detailed Description
The technical scheme of the invention is not limited to the specific embodiments listed below, and also includes any combination of the specific embodiments.
The first embodiment is as follows: the molecular structure of the fluorescent probe for relay recognition of zinc ions and glyphosate in the embodiment is as follows:
the second embodiment is as follows: the preparation method of the fluorescent probe comprises the following steps: reflux of the 2-hydroxy-1-naphthaldehyde and the 2-aminoquinoline in an ethanol solution, and condensation reaction to obtain the fluorescent probe.
And a third specific embodiment: the fluorescent probe of the embodiment is applied to relay recognition of zinc ions and glyphosate.
The specific embodiment IV is as follows: the third difference between this embodiment and the third embodiment is that: the operation method for identifying zinc ions and glyphosate by the fluorescent probe in relay mode comprises the following steps: in DMSO solution, the probe can detect zinc ion "turn-on", and after glyphosate is directly added into the detection system, the probe and zinc ion complex can detect glyphosate "on-off".
Fifth embodiment: this practice isThe fourth embodiment differs from the first embodiment in that: the detection limit of the fluorescent probe on zinc ions is 4.7X10 -7 The detection limit of the probe and zinc ion complex to glyphosate is 2.5X10 mol/L -7 mol/L. The other is the same as in the fourth embodiment.
Specific embodiment six: the present embodiment differs from the fifth or sixth embodiment in that: the fluorescent probe has high selectivity, high anti-interference performance, high sensitivity and visual relay identification on zinc ions and glyphosate. The other is the same as in the fifth or sixth embodiment.
Seventh embodiment: this embodiment differs from the fourth or fifth or sixth embodiment in that: the fluorescent probe can be used for reversible cyclic recognition of zinc ions and glyphosate. The others are the same as those of the fourth, fifth or sixth embodiments.
Eighth embodiment: the present embodiment differs from the fourth to seventh embodiments in that: the fluorescent probe forms a complex with zinc ions for detecting residues of glyphosate in soil. The others are the same as in the fourth to seventh embodiments.
When the glyphosate-containing sample is actually detected, the sample is subjected to pretreatment of centrifugation and filtration to remove solid particle impurities.
The following examples of the present invention are described in detail, and are provided by taking the technical scheme of the present invention as a premise, and the detailed embodiments and specific operation procedures are given, but the scope of the present invention is not limited to the following examples.
Example 1: the preparation method of the fluorescent probe comprises the following steps:
in a 100mL three-necked flask, 0.5g (2.9 mmol) of 2-hydroxy-1-naphthamaldehyde, 0.42g (2.9 mmol) of 2-aminoquinoline, a little glacial acetic acid and 50mL of ethanol were added and the mixture was refluxed for 6 hours. Suction filtration while hot, ethanol recrystallization and vacuum drying gave fluorescent probe L0.73g (2.4 mmol) in 84% yield. 1 H NMR(300MHz,DMSO-d 6 )δ15.90(d，J＝11.4Hz,1H)，9.61(d，J＝11.4Hz,1H),9.04(dd，J＝4.2，1.7Hz，1H)，8.45(td，J＝8.5，1.4Hz，3H)，7.93–7.77(m，2H)，7.77–7.59(m，3H)，7.50(ddd，J＝8.4，7.1，1.4Hz，1H)，7.37–7.13(m，1H)，6.72(d，J＝9.5Hz，1H). 13 C NMR(300MHz,DMSO-d 6 )δ181.82，150.63，147.74，139.83，139.21，136.90，136.75，134.69，129.57，128.87，128.82，127.42，126.73，126.39，124.91，123.92，123.04，120.38，115.11，108.71ppm.FT-IR(KBr)：3417.65，3057.82，1625.78，1611.34，1539.20，1472.00，1303.21，1209.43，1136.54，1083.13，958.79，820.71，789.13cm -1 .
Fluorescent probes 1 H NMR spectrum, 13 The C NMR spectrum and the IR spectrum are shown in FIGS. 2, 3 and 4, respectively.
Example 2: the fluorescent probe of the embodiment selectively recognizes zinc ions, and comprises the following steps:
cu with concentration of 0.1M was prepared 2+ 、Ni 2+ 、Co 2+ 、Cd 2+ 、Mg 2+ 、Pb 2+ 、Ca 2+ 、Ce 3+ 、Hg 2+ 、Zn 2+ 、Na + 、K + 、Fe 3+ 、Ag + 、Ba 2+ 、Cs + 、Al 3+ 、Cr 3+ The aqueous solution is ready for use.
Accurately weighing 8.9mg fluorescent probe and dissolving in 100mL DMSO solution to prepare 3.0X10 -4 mol/L solution A; taking 10mL of solution A, and fixing the volume to a 100mL volumetric flask by using DMSO solution to prepare 3.0X10 -5 And (5) mol/L of the solution B for standby.
3mL of solution B are taken in a cuvette at a time, 1eq. Of Cu is added in turn 2+ 、Ni 2+ 、Co 2+ 、Cd 2+ 、Mg 2+ 、Pb 2+ 、Ca 2+ 、Ce 3+ 、Hg 2+ 、Zn 2+ 、Na + 、K + 、Fe 3+ 、Ag + 、Ba 2+ 、Cs + 、Al 3+ 、Cr 3+ An aqueous solution. Under the action of 420nm exciting light, measuring the fluorescence emission peak intensity value of a fluorescent probe at 537nm, and adding Zn into the fluorescent probe 2+ After that, the fluorescent probe has obvious fluorescence enhancement effect, but the addition of other metal ions can not cause obvious fluorescence change, and the fluorescent probe is specific to Zn 2+ Exhibit specificity ofThe result of the recognition is shown in fig. 5.
Example 3: the detection limit of the fluorescent probe for zinc ions in the embodiment is carried out according to the following steps:
3mL of the solution was taken to have a concentration of 3.0X10 -5 1. Mu.L of solution B was added at a concentration of 3X 10 -3 The fluorescence intensity was measured from the mol/L zinc ion aqueous solution. The results are shown in FIG. 6, plotted with the zinc ion concentration on the abscissa and the fluorescence intensity on the ordinate.
As can be obtained by data analysis, when the zinc ion concentration is within 0 mu M-40 mu M, the fluorescence intensity is continuously enhanced along with the increase of the zinc ion concentration, the fluorescence intensity and the zinc ion concentration show good linear relation, the fitting equation is y=7.57x+47.66, and R is the formula 2 = 0.9902. According to the calculation formula 3 sigma/k of the detection limit, the detection limit of the probe molecule to the zinc ions is calculated to be 4.7x10 - 7 mol/L. The probe can realize trace detection of zinc ions.
Example 4: the complexing ratio of the fluorescent probe and zinc ions in the embodiment is carried out according to the following steps:
the concentration is 3.0X10 -5 mol/L solution B and 3X 10 -3 The mol/L zinc ion aqueous solution keeps the total degree of the probe and the zinc ion in the system to be 9.0x10 -8 The mol is unchanged, the fluorescence intensity is measured by changing the equivalent ratio of the probe and the zinc ion, and a Job's Plot is drawn. The results are shown in FIG. 7.
As can be seen from analysis of the Job's Plot, when the mole fraction of zinc ions was 0.56, the fluorescence intensity appeared to be inflection point, thereby indicating that the probe L and Zn 2+ The complexing ratio is 1:1.
Example 5: fluorescent probe-Zn 2+ The selective identification of glyphosate is carried out according to the following steps:
3mL of the solution was taken to have a concentration of 3.0X10 -5 Adding 1eq of Zn ion solution into mol/L solution B, incubating for 3min to obtain a complex L-zinc ion system, sequentially adding 1eq of glyphosate, trichlorfon, thioimine, dichlorvos, malathion, omethoate, dimethoate, acephate, fenitrothion, methyl parathion, parathion and glufosinate, measuring fluorescence intensity under the action of 420nm excitation light, and obtaining the result as shown in figure 8Shown.
According to data analysis, when glyphosate is added, the fluorescence of the system is obviously quenched, and when other pesticides are added, the fluorescence intensity is not obviously changed, and the fluorescence probe is Zn 2+ The system realizes the specific recognition of the glyphosate.
Example 6: fluorescent probe-Zn under ultraviolet light (365 nm) 2+ Systematic qualitative detection of glyphosate
1.5mL of fluorescent probe-Zn is accurately measured 2+ And 2eq. Of glyphosate, trichlorfon, thiotepa, dichlorvos, malathion, omethoate, dimethoate, acephate, fenitrothion, methyl parathion, parathion and glufosinate are respectively added into the system C, and the result shown in figure 9 is obtained under the ultraviolet light of 365 nm. As can be seen from FIG. 9, the fluorescent probe-Zn 2+ After the system is acted with organophosphorus pesticide, only glyphosate has obvious fluorescence change, (wherein 1# to 14# correspond to probes, and probes are Zn 2+ Glyphosate, trichlorfon, iminothiolane, dichlorvos, malathion, omethoate, dimethoate, acephate, fenitrothion, methyl parathion, parathion and glufosinate), shows excellent selectivity to glyphosate, can be used for qualitative detection of glyphosate, and has simple and convenient operation and high detection speed.
Example 7: fluorescent probe-Zn 2+ The glyphosate identification anti-interference performance is carried out according to the following steps:
3mL of the solution was taken to have a concentration of 3.0X10 -5 Adding 1eq. Zinc ion solution into mol/L solution B, and incubating for 3min to obtain fluorescent probe-Zn 2+ And (3) sequentially adding 1eq of glyphosate, trichlorfon, iminothiolane, dichlorvos, malathion, omethoate, dimethoate, cyphos, fenitrothion, methyl parathion, parathion and glufosinate into the system, recording the fluorescence intensity under the excitation light effect of 420nm, sequentially adding 1eq of glyphosate, and observing and recording the fluorescence intensity change. The results are shown in FIG. 10.
As can be obtained by data analysis, after the glyphosate is added, the fluorescence quenching can be realized after the glyphosate is added in the presence of other coexisting pesticides, and other pesticides can be used for fluorescence probe-Zn 2+ The system recognizes that glyphosate is interference free.
Example 8: fluorescent probe-Zn 2+ The detection limit of the glyphosate is carried out according to the following steps:
the concentration is 3.0X10 -5 Adding 1eq. Zinc ion solution into the mol/L solution B, and incubating for 3min to obtain fluorescent probe L-Zn 2+ The system is ready for use, 3mL fluorescent probe L-Zn is taken 2+ The system was added at a concentration of 3X 10 in 1. Mu.L each time -3 The fluorescence intensity was measured from the aqueous solution of mol/L glyphosate. The results are shown in FIG. 11.
As can be seen from data analysis, when the concentration of the glyphosate is within 0 mu M-24 mu M (namely 0-4 mu g/mL), the fluorescence intensity is continuously weakened along with the increase of the concentration of the glyphosate, the fluorescence intensity and the concentration of the glyphosate show good linear relation, and a fitting equation is that y= 326.96-10.69X and R 2 = 0.9979. Calculating fluorescent probe-Zn according to the calculation formula 3 sigma/k of the detection limit 2+ The detection limit of glyphosate ion is 2.5 multiplied by 10 -7 mol/L (i.e., 42.26 ng/mL). Fluorescent probe-Zn 2+ The system can realize trace detection of glyphosate.
Example 9: fluorescent probe-Zn 2+ The glyphosate action ratio is carried out according to the following steps:
fluorescent probe-Zn 2+ System and 3X 10 -3 mol/L glyphosate aqueous solution, and maintaining fluorescent probe-Zn in system 2+ The total degree of system and glyphosate is 9x 10 -8 The mol is unchanged by changing the fluorescent probe-Zn 2+ The equivalent ratio of the system to glyphosate was measured for fluorescence intensity and a Plot of Job's Plot was drawn. The results are shown in FIG. 12.
As can be seen from the data analysis, when the mole fraction of glyphosate was 0.45, the fluorescence intensity appeared to be inflection point, thus indicating that the fluorescent probe-Zn 2+ The complexing ratio of the system and the glyphosate is 1:1.
Example 10: fluorescent probe pair Zn 2+ And reversible cycling of glyphosate
In order to further explore the recognition characteristics of the probe on zinc ions and glyphosate, 3mL of probe solution is taken, zinc ions and glyphosate solution are alternately added, fluorescence intensity is recorded, and a circulation curve is drawn. The results are shown in FIG. 13.
Sensor for detecting a position of a bodyAlternate identification of Zn 2+ And in the process of glyphosate, the fluorescence intensity breakage rate is about 5% when the glyphosate is continuously circulated for 5 times, so that the sensor can realize the alternate fluorescence identification of glyphosate and zinc ions. Meanwhile, the mechanism of the sensor for identifying the glyphosate is further proved to be that the glyphosate can chelate zinc ions in a sensing system, so that the sensor is restored to the initial state of the probe.
Example 11: fluorescent sensor for detecting glyphosate in actual soil
In order to examine the potential application of the fluorescent probe in the actual environment, field soil is selected, and the soil is pretreated: weighing 1g of soil, placing into a centrifuge tube, adding 50mL of water, ultrasonically extracting for 30min, centrifuging for 5min at 8000r/min, filtering supernatant with filter paper to obtain soil extract, and preparing glyphosate solutions with concentrations of 0.5 mug/mL, 1.0 mug/mL, 2.0 mug/mL and 3.0 mug/mL respectively
Fluorescent probe L-Zn prepared by example 6 2+ The system is to fluorescent probe L-Zn 2+ Adding the glyphosate solutions with different concentrations into the system, and measuring the fluorescent probe L-Zn under the excitation light effect of 420nm 2+ The fluorescence emission peak intensity value of the system at 537nm is brought into the following equation, and the concentration of the glyphosate solution to be detected is calculated. The test results are shown in Table 1.
Y＝326.96-10.69X
Wherein X is the concentration of glyphosate, and Y is the fluorescence emission peak intensity value.
TABLE 1 fluorescent Probe-Zn 2+ System for detecting glyphosate in actual water sample
As can be seen from Table 1, the recovery rate of glyphosate in actual soil is 98.2% -105.5%, the relative standard deviation is 3.07%, and the measured concentration of glyphosate has small error with the correspondent standard-added concentration, and these results indicate that the fluorescent probe-Zn prepared by the invention 2+ The system has better accuracy in detecting the glyphosate in the actual soil, and can be in the range of 0 to ultraThe glyphosate is quantitatively detected within the range of 4.0 mug/mL, and the method has good practical performance.
Claims (8)
1. A fluorescent probe for identifying zinc ions and glyphosate in a relay manner is characterized in that the molecular structure of the fluorescent probe is as follows:
。
2. the method for preparing the fluorescent probe for identifying zinc ions and glyphosate in relay according to claim 1, which is characterized by comprising the following steps: reflux of the 2-hydroxy-1-naphthaldehyde and the 2-aminoquinoline in an ethanol solution, and condensation reaction to obtain the fluorescent probe.
3. Use of the fluorescent probe of claim 1 for the relay recognition of zinc ions and glyphosate.
4. The use according to claim 3, wherein the fluorescent probe is operative to identify zinc ions and glyphosate in relay as: in DMSO solution, the probe can detect zinc ion "turn-on", and after glyphosate is directly added into the detection system, the probe and zinc ion complex can detect glyphosate "on-off".
5. The use of the fluorescent probe for the relay recognition of zinc ions and glyphosate as claimed in claim 4, wherein the detection limit of the fluorescent probe on zinc ions is 4.7X10 -7 The detection limit of the probe and zinc ion complex to glyphosate is 2.5X10 mol/L -7 mol/L。
6. The use of the fluorescent probe according to claim 4 for relay identification of zinc ions and glyphosate, wherein the fluorescent probe can realize relay identification with high selectivity, high anti-interference performance, high sensitivity and visualization of zinc ions and glyphosate.
7. The use of the fluorescent probe according to claim 4 for the relay recognition of zinc ions and glyphosate, wherein the fluorescent probe is capable of achieving reversible cycle recognition of zinc ions and glyphosate.
8. The use of the fluorescent probe according to claim 4 for identifying zinc ions and glyphosate in relay, wherein the complex formed by the fluorescent probe and zinc ions can be used for detecting glyphosate residues in soil.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107098857A (en) * | 2017-06-13 | 2017-08-29 | 黄淮学院 | Fluorescence probe for recognizing zinc ion and its preparation method and application, membrane material for recognizing zinc ion and preparation method thereof |
| CN109970634A (en) * | 2019-03-27 | 2019-07-05 | 福建师范大学 | A kind of pyridine schiff base structure fluorescence probe and the preparation method and application thereof for detecting copper ion |
| CN113788789A (en) * | 2021-10-13 | 2021-12-14 | 哈尔滨理工大学 | Preparation method and application of fluorescent probe for continuously detecting copper ions and glyphosate |
| CN113861147A (en) * | 2021-10-08 | 2021-12-31 | 哈尔滨理工大学 | Preparation method and application of fluorescent sensor for detecting glyphosate |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110117282B (en) * | 2019-05-13 | 2020-07-10 | 五邑大学 | Zinc ion fluorescent probe compound and preparation method and application thereof |
-
2022
- 2022-04-04 CN CN202210350350.5A patent/CN114835642B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107098857A (en) * | 2017-06-13 | 2017-08-29 | 黄淮学院 | Fluorescence probe for recognizing zinc ion and its preparation method and application, membrane material for recognizing zinc ion and preparation method thereof |
| CN109970634A (en) * | 2019-03-27 | 2019-07-05 | 福建师范大学 | A kind of pyridine schiff base structure fluorescence probe and the preparation method and application thereof for detecting copper ion |
| CN113861147A (en) * | 2021-10-08 | 2021-12-31 | 哈尔滨理工大学 | Preparation method and application of fluorescent sensor for detecting glyphosate |
| CN113788789A (en) * | 2021-10-13 | 2021-12-14 | 哈尔滨理工大学 | Preparation method and application of fluorescent probe for continuously detecting copper ions and glyphosate |
Non-Patent Citations (5)
| Title |
|---|
| Luminescent property and catalytic activity of Ru(II) carbonyl complexes containing N, O donor of 2-hydroxy-1-naphthylideneimines;M. Sivagamasundari等;《Spectrochimica Acta Part A》;第67卷(第1期);第427-433页 * |
| NPA-Cu2+ Complex as a Fluorescent Sensing Platform for the Selective and Sensitive Detection of Glyphosate;Fang Sun等;《International Journal of Molecular Sciences》;第22卷(第18期);第9816页 * |
| 一种具有聚集荧光增强性质苯并咪唑化合物的合成及其对Zn2+的识别;边延江等;《化学研究与应用》;第31卷(第5期);第819-823页 * |
| 一种基于8-氨基喹啉的希夫碱型荧光探针对锌离子的荧光识别;汤立军等;《渤海大学学报（自然科学版）》;第37卷(第4期);第330-335页 * |
| 基于喹啉功能团的Zn2+荧光探针;吴红梅等;《发光学报》;第29卷(第6期);第1091-1095页 * |
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