CN116574062A - Fluorescent probe for naked eye and real-time detection of hypochlorite, and preparation method and application thereof - Google Patents
Fluorescent probe for naked eye and real-time detection of hypochlorite, and preparation method and application thereof Download PDFInfo
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- CN116574062A CN116574062A CN202310531233.3A CN202310531233A CN116574062A CN 116574062 A CN116574062 A CN 116574062A CN 202310531233 A CN202310531233 A CN 202310531233A CN 116574062 A CN116574062 A CN 116574062A
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- WQYVRQLZKVEZGA-UHFFFAOYSA-N hypochlorite Chemical compound Cl[O-] WQYVRQLZKVEZGA-UHFFFAOYSA-N 0.000 title claims abstract description 171
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 108
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 238000011897 real-time detection Methods 0.000 title claims description 40
- 239000000523 sample Substances 0.000 claims abstract description 49
- 238000001514 detection method Methods 0.000 claims abstract description 33
- -1 hypochlorite ions Chemical class 0.000 claims abstract description 7
- 239000000243 solution Substances 0.000 claims description 79
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 46
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 36
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 30
- 238000006243 chemical reaction Methods 0.000 claims description 28
- 239000007787 solid Substances 0.000 claims description 27
- 239000012085 test solution Substances 0.000 claims description 27
- 239000012043 crude product Substances 0.000 claims description 25
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 24
- 239000003960 organic solvent Substances 0.000 claims description 23
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 21
- GNKZMNRKLCTJAY-UHFFFAOYSA-N 4'-Methylacetophenone Chemical compound CC(=O)C1=CC=C(C)C=C1 GNKZMNRKLCTJAY-UHFFFAOYSA-N 0.000 claims description 20
- 238000010992 reflux Methods 0.000 claims description 19
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 17
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- 239000011550 stock solution Substances 0.000 claims description 14
- 238000001914 filtration Methods 0.000 claims description 13
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 12
- 229960000583 acetic acid Drugs 0.000 claims description 12
- 230000005284 excitation Effects 0.000 claims description 12
- 238000002189 fluorescence spectrum Methods 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 12
- 239000007810 chemical reaction solvent Substances 0.000 claims description 11
- 239000012153 distilled water Substances 0.000 claims description 11
- 230000007935 neutral effect Effects 0.000 claims description 11
- 230000001105 regulatory effect Effects 0.000 claims description 11
- 238000003756 stirring Methods 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 238000010438 heat treatment Methods 0.000 claims description 10
- 238000005303 weighing Methods 0.000 claims description 10
- 239000008055 phosphate buffer solution Substances 0.000 claims description 9
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 8
- 238000009835 boiling Methods 0.000 claims description 8
- 238000001816 cooling Methods 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 8
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 6
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 6
- 238000007865 diluting Methods 0.000 claims description 6
- 239000012362 glacial acetic acid Substances 0.000 claims description 6
- 239000012488 sample solution Substances 0.000 claims description 6
- YYVYAPXYZVYDHN-UHFFFAOYSA-N 9,10-phenanthroquinone Chemical compound C1=CC=C2C(=O)C(=O)C3=CC=CC=C3C2=C1 YYVYAPXYZVYDHN-UHFFFAOYSA-N 0.000 claims description 5
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 claims description 5
- 239000005695 Ammonium acetate Substances 0.000 claims description 5
- 229940043376 ammonium acetate Drugs 0.000 claims description 5
- 235000019257 ammonium acetate Nutrition 0.000 claims description 5
- 239000012065 filter cake Substances 0.000 claims description 5
- 238000005259 measurement Methods 0.000 claims description 5
- 239000000126 substance Substances 0.000 claims description 5
- KUCOHFSKRZZVRO-UHFFFAOYSA-N terephthalaldehyde Chemical compound O=CC1=CC=C(C=O)C=C1 KUCOHFSKRZZVRO-UHFFFAOYSA-N 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 4
- MVPPADPHJFYWMZ-UHFFFAOYSA-N chlorobenzene Chemical compound ClC1=CC=CC=C1 MVPPADPHJFYWMZ-UHFFFAOYSA-N 0.000 claims description 4
- 239000000706 filtrate Substances 0.000 claims description 4
- 239000002904 solvent Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 3
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 claims description 3
- 238000000967 suction filtration Methods 0.000 claims description 3
- 239000006227 byproduct Substances 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 230000001376 precipitating effect Effects 0.000 claims description 2
- 238000011084 recovery Methods 0.000 claims description 2
- 230000007613 environmental effect Effects 0.000 abstract 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 12
- 239000007788 liquid Substances 0.000 description 12
- 230000008859 change Effects 0.000 description 6
- 239000011780 sodium chloride Substances 0.000 description 6
- 238000001228 spectrum Methods 0.000 description 6
- 150000001450 anions Chemical class 0.000 description 5
- 238000005481 NMR spectroscopy Methods 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 239000011259 mixed solution Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 239000001257 hydrogen Substances 0.000 description 3
- 238000010791 quenching Methods 0.000 description 3
- 230000000171 quenching effect Effects 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- IOJUPLGTWVMSFF-UHFFFAOYSA-N benzothiazole Chemical class C1=CC=C2SC=NC2=C1 IOJUPLGTWVMSFF-UHFFFAOYSA-N 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000002795 fluorescence method Methods 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 230000001590 oxidative effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- 102000003896 Myeloperoxidases Human genes 0.000 description 1
- 108090000235 Myeloperoxidases Proteins 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 1
- 238000000295 emission spectrum Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000005281 excited state Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000001506 fluorescence spectroscopy Methods 0.000 description 1
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 125000001484 phenothiazinyl group Chemical class C1(=CC=CC=2SC3=CC=CC=C3NC12)* 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 229940066767 systemic antihistamines phenothiazine derivative Drugs 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D235/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings
- C07D235/02—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, condensed with other rings condensed with carbocyclic rings or ring 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
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6447—Fluorescence; Phosphorescence by visual observation
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1044—Heterocyclic compounds characterised by ligands containing two nitrogen atoms as heteroatoms
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Life Sciences & Earth Sciences (AREA)
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- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Materials Engineering (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Optics & Photonics (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
A fluorescent probe for detecting hypochlorite in real time with naked eyes and a preparation method and application thereof relate to a fluorescent probe for detecting hypochlorite and a preparation method and application thereof. It aims to solve the existing ClO – The probe can not realize the technical problems of naked eye detection, harsh use environment, poor recognition specificity and higher detection limit. The structural formula of the fluorescent probe for detecting hypochlorite in real time and naked eyes of the invention is as follows:it uses 4 (1-H-phenanthro [9, 10-d)]The fluorescent probe is used for qualitative or quantitative detection of hypochlorite ions in a solution, can be identified by naked eyes and detected in real time, and is not affected by the fact that the fluorescent probe is synthesized by imidazole-2) -benzaldehyde and 4-methylacetophenoneThe influence of other interferents in the environment can be used in the environment with pH of 4-12, clO – The detection limit is as low as 46.19nmol/L. ClO useful in environmental and biological systems – Qualitative and quantitative detection of (a).
Description
Technical Field
The invention relates to a fluorescent probe for detecting hypochlorite, a preparation method and application thereof.
Background
Hypochlorous acid in its protonated form is the most important active oxygen species in the organism, usually produced by the reaction of hydrogen peroxide with chloride ions, and the reaction is catalyzed by myeloperoxidase. Compared with the traditional hypochlorite detection method, the fluorescence method for detecting hypochlorite is used as a non-invasive detection technology, and scientific research is widely focused by virtue of numerous advantages (such as high sensitivity, good selectivity, rapid response, in-situ detection, real-time monitoring and the like).
In recent years, many researchers have developed research work on the design and application of hypochlorite ion fluorescent probes. In 2023, liu Meng et al report on pages 345-351 of volume 34 of organic chemistry on page 12 that a benzothiazole derivative fluorescent probe based on the mechanism of aggregation-induced emission and intramolecular proton transfer in excited state and its recognition of hypochlorite synthesized a benzothiazole derivative-based ClO – Fluorescent probe, the probe pair ClO – Has better recognition capability along with ClO – The concentration is increased, and the cells gradually change from non-fluorescence to red fluorescence under the irradiation of an ultraviolet lamp, but naked eye detection cannot be realized. In 2017, li Hongyu et al, analytical chemistry, vol.51, pp.5, 619-626, use sensitive fluorescent probes to observe ONOO in mitochondria under various stimuli – Is reported in (1) a hydrazide oxidation-based hypochlorite probe that is specific for ClO – Poor recognition specificity, mainly in response to ClO – Is also susceptible to oxidative species such as nitrosylperoxideInterference. In 2020, wang Xiaolin et al, vol.37, vol.2, pp.93-105, organic chemistry, synthesis of phenothiazine-rhodanine fluorescent probes for ClO, et al, synthesis of fluorescent probes based on phenothiazine derivatives for use in cell imaging applications – Has good selectivity, but the probe has the same effect on ClO – Is of the detection limit (94.18 nmol.L) –1 ) Higher, not suitable for biological detection.
According to the current literature report, clO – The probe has mainly the following defects:
1. most ClO – The fluorescent probe cannot realize naked eye real-time detection;
2. the identification specificity is poor, and the interference of other oxidation species is easy to occur;
3. the detection limit is higher, and the application range is limited.
Thus, clO with in-situ real-time detection, high specificity, high sensitivity and easy operation is designed and developed – The fluorescent probe has important practical significance.
Disclosure of Invention
The invention aims to solve the problems of the existing ClO – The fluorescent probe for detecting hypochlorite in real time and the preparation method and application thereof can realize naked eye detection, has mild use environment, special recognition, low detection limit and easy operation.
The structural formula of the fluorescent probe for detecting hypochlorite in real time and naked eyes of the invention is as follows:
the preparation method of the fluorescent probe for naked eyes and real-time detection of hypochlorite is carried out according to the following steps:
1. the ratio of the amounts of the substances of 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde to 4-methylacetophenone was 1: (1-6) respectively weighing 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde v and 4-methylacetophenone, adding the mixture into a three-mouth bottle with a reflux device, adding an organic solvent I as a reaction solvent, heating to boiling and refluxing under stirring, keeping for 1-10 hours for reaction, cooling to room temperature after the reaction is finished, filtering to remove solid byproducts, collecting filtrate, adding distilled water into the filtrate, regulating the pH value to be neutral by using a 5% NaOH solution, precipitating solids, filtering, collecting filter cakes, and obtaining a crude product;
2. recrystallizing the crude product by using an organic solvent II to obtain a fluorescent probe for detecting hypochlorite in real time with naked eyes, wherein the fluorescent probe is a dark yellow solid.
Further, the organic solvent I in the first step is methanol, ethanol, ethyl acetate, acetonitrile, dichloromethane, glacial acetic acid or acetone.
Further, the organic solvent II in the third step is ethanol, methanol, propanol, chloroform, dichloromethane or ethyl acetate.
Further, the preparation method of the 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde in the step one comprises the following steps:
(1) The ratio of the amounts of the substances is 1: (1-3): (1-5) weighing phenanthrenequinone, terephthalaldehyde and ammonium acetate, adding the phenanthrenequinone, the terephthalaldehyde and the ammonium acetate into an organic solvent A as a reaction solvent, stirring at room temperature for 5-20 hours, and stopping the reaction; wherein the organic solvent A is benzene, chlorobenzene, glacial acetic acid or N, N-dimethylformamide;
(2) Distilled water is added into the reaction system, the pH value of the reaction system is regulated to be neutral by using a NaOH solution with the concentration of 0.1mol/L, solids are separated out, suction filtration is carried out, and a filter cake is collected, so that a crude product is obtained;
(3) Adding the crude product into an organic solvent B, heating to the reflux temperature of the solvent, recrystallizing, filtering while the crude product is hot, and drying to obtain 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde; wherein the organic solvent B is ethyl acetate, dichloromethane, chloroform, acetone, ethanol or methanol.
The synthesis of 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde according to the present invention can be represented by the following formula:
the synthesis of the fluorescent probe for naked eye and real-time hypochlorite detection of the present invention can be represented by the following formula:
the application of the fluorescent probe for detecting hypochlorite ions in real time and with naked eyes is to use the fluorescent probe for qualitative or quantitative detection of hypochlorite ions in solution.
ClO in aqueous solution by using naked eyes and fluorescent probe pair for detecting hypochlorite in real time – The naked eye qualitative detection method comprises the following steps:
1. dissolving a fluorescent probe for detecting hypochlorite in real time with naked eyes in an organic solvent to prepare a probe stock solution A;
2. diluting the probe stock solution A obtained in the step one to a concentration of 0.01-0.05 mmol.L by using PBS phosphate buffer solution –1 A probe solution B;
3. adding a solution to be tested into the probe solution B in the second step to obtain a test solution C;
4. after the test solution C stands for 2-3 min, observing the color of the test solution C under natural light, and judging that the solution to be tested contains ClO if the color is changed from yellow to colorless – The method comprises the steps of carrying out a first treatment on the surface of the Or after the test solution C stands for 2-3 min, observing the color of the test solution C under the irradiation of a handheld ultraviolet lamp, and judging that the solution to be tested contains ClO if the color is changed from pink to green – 。
Further, the organic solvent in the first step is ethyl acetate, methanol, ethanol, glacial acetic acid, acetone or acetonitrile.
Further, the concentration of the fluorescent probe for detecting hypochlorite in real time with naked eyes in the first step is 0.1 mmol.L –1 。
Further, the concentration of the PBS phosphate buffer solution in the second step is 0.01mol·L –1 ，pH＝7.40。
Further, the wavelength of the hand-held ultraviolet lamp in the fifth step is 365nm.
ClO in solution by using naked eyes and real-time detection hypochlorite fluorescent probe – The fluorescence qualitative detection method of (2) comprises the following steps:
1. dissolving a fluorescent probe for detecting hypochlorite in real time with naked eyes in an organic solvent to prepare a probe stock solution A;
2. diluting the probe stock solution A obtained in the step one to a concentration of 0.01-0.05 mmol.L by using PBS phosphate buffer solution –1 A probe solution B;
3. measuring fluorescence emission spectrum of probe solution B with 420nm as excitation wavelength, recording emission intensity at 615nm, and recording as T B ；
4. Adding a solution to be tested into the probe solution B in the second step to obtain a test solution C;
5. after the test solution C is kept stand for 2 to 3 minutes, the fluorescence emission spectrum of the test solution C is measured by taking 420nm as an excitation wavelength, and the emission intensity when the emission wavelength is 615nm is recorded and is recorded as T C ；
6. Comparison T B And T C If 4T B ≤T C Judging that the solution to be tested contains ClO – 。
ClO in solution by using naked eyes and real-time detection hypochlorite fluorescent probe – The fluorescent quantitative detection method of (2) is a standard curve method, and is specifically carried out according to the following steps:
1. dissolving a fluorescent probe for detecting hypochlorite in real time with naked eyes in an organic solvent to prepare a probe stock solution A;
2. diluting the probe stock solution A obtained in the step one to a concentration of 0.01-0.05 mmol.L by using PBS phosphate buffer solution –1 A probe solution B;
3. gradually adding the probe solution B to the probe solution B at the concentration of C respectively 1 、C 2 、C 3 ……C n ClO of (C) – Solutions, respectively obtaining sample solutions B 1 、B 2 、B 3 ……B n ；
4. Measurement of sample solution B with excitation wavelength of 420nm 1 ～B n The fluorescence emission spectra of (2) were recorded as T, and the emission intensity at 615nm was recorded, respectively B1 ～T Bn ；
5. In ClO – Concentration C of (2) 1 ～C n On the abscissa, with the emission intensity T of the sample solution B1 ～T Bn Plotted as ordinate, clO – A standard curve between concentration-fluorescence intensity;
6. adding the solution to be tested into the probe solution B, and uniformly mixing to prepare a test solution B x ；
7. 2.0. Mu.M ClO was added to the test solution Bx – Measuring fluorescence emission spectrum of the test solution Bx by using 420nm as excitation wavelength, recording emission intensity at 615nm, and recording as T Bx ；
8. Will T Bx Looking up T by comparing the ordinate of the standard curve in the fifth step Bx ClO in standard curve of intensity correspondence – Determining ClO in the solution to be tested according to the recovery rate and average deviation after 3 times of measurement – Concentration.
The fluorescent probe for detecting hypochlorite in real time and naked eyes belongs to a reactive probe, wherein the oxidizing property of hypochlorite is utilized to oxidize C=C double bonds in the probe to break C=C, the structure of the probe is changed, and then blue shift of fluorescence emission wavelength and quenching of fluorescence intensity are caused. The fluorescent probe pair ClO – The fluorescent probe has stronger identification specificity and real-time monitoring capability, can realize naked eye and real-time detection, is not influenced by other interferents in the environment, can be used in a wider environment with pH value of 4-12, and changes in fluorescence intensity and ClO of the fluorescent probe – The change in concentration corresponds to a linear relationship, clO – The detection limit is as low as 46.19nmol/L.
The fluorescent probe for naked eyes and real-time detection of hypochlorite is simple in synthesis method, low in cost, simple in steps, mild in reaction condition and high in yield. Detection of ClO – The method is simple, has obvious effect and quick response, and can be used for ClO in environment and biological systems through visual effect or fluorescence spectrum – Qualitative and quantitative detection of (a).
Drawings
FIG. 1 is a hydrogen nuclear magnetic spectrum of 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde prepared in step one of example 1;
FIG. 2 is a carbon nuclear magnetic spectrum of 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde prepared in step one of example 1;
FIG. 3 is a sample of the fluorescent probe solution for naked-eye and real-time detection of hypochlorite prepared in example 1 (10. Mu. Mol.L) –1 ，V Ethanol :V PBS =8:2, ph=7.40) in the presence of different types of oxides (10 μmol·l –1 ) Fluorescent emission spectrum of (lambda) ex =420 nm), the abscissa is wavelength, and the ordinate is fluorescence intensity;
FIG. 4 is a sample of the fluorescent probe solution for naked-eye and real-time detection of hypochlorite prepared in example 1 (10. Mu. Mol.L) –1 ，V Ethanol :V PBS =8:2, ph=7.40) in other interfering species (20 μmol·l –1 ) Adding ClO in the presence of – Post fluorescence intensity histogram, abscissa is interference species (F – 、Cl – 、Br – 、I – 、S 2– 、BrO 3 – 、SO 3 2– 、SO 4 2– 、HPO 4 2– 、H 2 PO 4 – 、NO 2 – 、HSO 3 – 、NO 3 – 、AcO – 、CO 3 2– 、SCN – And HCO 3 – ) The ordinate is fluorescence intensity;
FIG. 5 is a sample of the fluorescent probe solution for naked-eye and real-time detection of hypochlorite prepared in example 1 (10. Mu. Mol.L) –1 ，V Ethanol :V PBS =8:2, ph=7.40) under different pH conditions, the abscissa is pH value, and the ordinate is fluorescence intensity;
FIG. 6 is an open-hole and real-time inspection prepared in example 1Fluorescent probe solution for measuring hypochlorite (10. Mu. Mol.L) –1 ) In ClO – (10μmol·L –1 ) A graph of fluorescence intensity over time in the presence, with time on the abscissa and fluorescence intensity on the ordinate;
FIG. 7 is a graph showing the fluorescence intensity and ClO of the fluorescent probe solution for bare eye and real-time detection of hypochlorite prepared in example 1 – Standard graph of concentration.
Detailed Description
The following examples are used to demonstrate the benefits of the present invention.
Example 1: the preparation method of the fluorescent probe for detecting hypochlorite in real time with naked eyes in the embodiment comprises the following steps:
1. weighing 0.32g of 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde and 0.13mL of 4-methylacetophenone, adding into a three-mouth bottle with a reflux device, adding 60mL of ethanol as a reaction solvent, heating to boiling and refluxing under stirring, and keeping for 3 hours for reaction; cooling to room temperature after the reaction is finished to obtain yellow liquid;
2. adding distilled water into the yellow liquid, regulating the pH value to be neutral by acetic acid, adding NaCl solution with the mass percent concentration of 20%, standing, and separating out solids; filtering out the solid, and drying to obtain a crude product;
3. the crude product was recrystallized from ethyl acetate to give a fluorescent probe for naked eye and real-time detection of hypochlorite, which was a dark yellow solid.
The preparation method of the 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde in the step one comprises the following steps:
(1) 1.04g (5.0 mmol) of phenanthrenequinone, 0.67g (5.0 mmol) of terephthalaldehyde and 0.39g (5.0 mmol) of ammonium acetate are respectively weighed, 20mL of glacial acetic acid is added as a reaction solvent, and the reaction is stopped after stirring for 10 hours at room temperature;
(2) Distilled water is added into the reaction system, the pH value of the reaction system is regulated to be neutral by using a NaOH solution with the concentration of 0.1mol/L, a large amount of solids are separated out, suction filtration is carried out, and a filter cake is collected, so that a crude product is obtained;
(3) The crude product is added into ethyl acetate and heated to a reflux temperature of 85 ℃ for recrystallization, and after filtration while hot, the crude product is dried to obtain 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde. The yield of 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde was 82%. The nuclear magnetic spectrum of the 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde hydrogen is shown in figure 1, and the nuclear magnetic spectrum of the carbon is shown in figure 2:
1 H NMR(600MHz,DMSO-d 6 )δ:13.740(s,1H,N-H),10.106(s,1H,CHO),8.885(d,J＝7.5Hz,2H,ArH),8.607(d,J＝6.7Hz,2H,ArH),8.547(d,J＝8.0Hz,2H,ArH),8.154(d,J＝8.1Hz,2H,ArH),7.772(s,2H,ArH),7.678(t,J＝7.4Hz,2H,ArH).
13 C NMR(150MHz,DMSO-d 6 )δ:193.00,192.97,175.09,149.91,137.70,135.85,130.50,128.09,127.39,126.90,126.02,125.43,124.26,122.69.ESI-MS(m/z)calcd.for(C 22 H 14 N 2 O 2 ,[M+H] + ):323.1184；found:323.1173.
characterization of the nuclear magnetic spectrum shows that the synthesized product is 4 (1-H-phenanthro [9, 10-d)]Imidazole-2) -benzaldehyde having the structure
The yield of the fluorescent probe for detecting hypochlorite ions with naked eyes and in real time prepared in the embodiment is 78%, and the melting point is more than 300 ℃.
The structure of the fluorescent probe for detecting hypochlorite in real time with naked eyes prepared in the embodiment is characterized by infrared spectrum and nuclear magnetic resonance hydrogen spectrum, and the following results are obtained:
1 H NMR(600MHz,DMSO-d 6 ):δ13.59(s,1H,NH),8.88(s,2H,ArH),8.61(s,3H,ArH),8.41(d,J＝7.9Hz,2H,ArH),8.16–8.11(m,2H,ArH),8.07(d,J＝15.5Hz,1H,CH),7.83(d,J＝15.6Hz,1H,CH),7.77(s,3H,ArH),7.66(d,J＝7.6Hz,2H,ArH),7.42(d,J＝7.8Hz,2H,ArH),2.39(s,3H,CH 3 ). As can be seen from the above structural characterization results, the naked eye and real-time hypochlorite fluorescent probe prepared in this embodiment has the structural formula:
ClO in aqueous solution using naked eye prepared in example 1 and fluorescent probe pair for real-time detection of hypochlorite – The naked eye qualitative detection is carried out, and the specific method comprises the following steps:
1. dissolving fluorescent probe for detecting hypochlorite in real time with naked eyes in ethanol to prepare the fluorescent probe with concentration of 0.1 mmol.L –1 Probe stock solution a of (a);
2. the probe stock solution A obtained in the first step was used at a concentration of 0.01 mol.L –1 PBS phosphate buffer solution with pH=7.40 is diluted to a concentration of 0.01 mmol.L –1 Is a probe solution B;
3. adding a solution to be detected into the probe solution B in the second step, wherein the solution to be detected contains ClO – Obtaining a test solution C;
4. after the test solution C is kept stand for 2-3 min, the color of the test solution C is observed under natural light, the color of the solution is changed from yellow to colorless, and the qualitative detection of ClO in the solution by observing the color change through naked eyes is shown – ；
After the test solution C is kept stand for 2 to 3 minutes, the color of the test solution C is observed under the irradiation of a handheld ultraviolet lamp, the color of the solution is changed from pink to green, and the fact that ClO in the solution can be qualitatively detected by a fluorescence method is shown – 。
The spectral properties of the naked eye and real-time hypochlorite fluorescent probe prepared in example 1 were tested by fluorescence spectrometry, and the steps are as follows:
1. the volume ratio of PBS phosphate buffer solution to ethanol is 2:8 (PBS buffer solution concentration of 0.01mol/L, pH=7.4) as solvent, and the fluorescent probe for naked eyes and real-time detection of hypochlorite prepared in example 1 was prepared to have a concentration of 1.0X10 －5 mol/L fluorescent probe solution;
2. respectively adding anions F to the fluorescent probe solution – 、Cl – 、Br – 、I – 、S 2– 、BrO 3 – 、SO 3 2– 、SO 4 2– 、HPO 4 2– 、H 2 PO 4 – 、NO 2 – 、HSO 3 – 、NO 3 – 、AcO – 、CO 3 2– 、SCN – And HCO 3 – The concentration of anions is 5.0X10 –4 mol/L to obtain the solution to be measured. In the solution to be detected, the mass ratio of the fluorescent probe of hypochlorite to the substance of anion is 1:50 by naked eyes and in real time;
3. after the solutions to be measured were left at room temperature for 5 minutes, the fluorescence spectra of the respective solutions to be measured were measured with an excitation wavelength of 420nm and an excitation slit width of 10.0nm, and the results are shown in FIG. 3.
The fluorescence emission wavelength of the fluorescent probe solution for bare-eye and real-time detection of hypochlorite prepared in example 1 was 615nm, and the fluorescence intensity was 1000a.u. After addition of the different anions, it can be seen that F – 、Cl – 、Br – 、I – 、S 2– 、BrO 3 – 、SO 3 2– 、SO 4 2– 、HPO 4 2– 、H 2 PO 4 – 、NO 2 – 、HSO 3 – 、NO 3 – 、AcO – 、CO 3 2– 、SCN – And HCO 3 – The effect on the fluorescence intensity of the fluorescent probe solution is not great, and the intensity is about 700a.u. To add ClO – After that, the emission wavelength was blue shifted to about 500nm, and the fluorescence intensity was quenched to about 290a.u., with a fluorescence intensity quenching ratio of 2.4 times. Thus, from the fluorescence emission spectrum, it can be determined that the fluorescent probe solution pair ClO – Having selective identification characteristics.
For the naked eye and real-time detection of hypochlorite fluorescent probe pair ClO prepared in this example 1 – The anti-interference capability of fluorescent identification is tested, and the specific method is as follows: :
1. the naked eye and real-time hypochlorite fluorescent probe prepared in example 1 was prepared to have a concentration of 1.0X10 s by using a mixed solution of PBS buffer and ethanol (wherein the concentration of PBS buffer is 0.01mol/L, pH=7.4) as a solvent –5 mol/L fluorescent probe solution. The anions are added into the fluorescent probe solution respectively to be F – 、Cl – 、Br – 、I – 、S 2– 、BrO 3 – 、SO 3 2– 、SO 4 2– 、HPO 4 2– 、H 2 PO 4 – 、NO 2 – 、HSO 3 – 、NO 3 – 、AcO – 、CO 3 2– 、SCN – And HCO 3 – Anion concentration 5.0X10 –4 And (3) mol/L, and uniformly mixing to obtain a mixed solution. Standing the mixed solution for 5min, and adding 5.0X10 g of the mixed solution respectively – 4 ClO of mol/L – The solution is mixed uniformly. Fluorescent probe/interferent/ClO at this time – The mass ratio of the three materials is 1:50:50. Measurement of fluorescent Probe/interferent/ClO with excitation wavelength at 420nm and excitation slit width at 10.0nm – The fluorescence emission spectrum and the result are shown in FIG. 4. As can be seen from fig. 4, clO – When coexisting with other interferents, the fluorescent intensity and ClO of hypochlorite fluorescent probe are detected in naked eyes and real time – The fluorescent intensity of the probe is not affected by the existence of other interferents when the probe exists alone, and the fluorescent probe pair ClO for naked eyes and real-time detection of hypochlorite – Has stronger anti-interference capability.
Under the condition of different pH values, detecting ClO by using fluorescent probe for detecting hypochlorite in naked eyes and real time – The trend of fluorescence intensity at 610nm in the front and rear is shown in FIG. 5. As can be seen from FIG. 5, the fluorescence intensity of the fluorescent probe for detecting hypochlorite in naked eyes and in real time is basically unchanged in a wide pH value range, which indicates that the fluorescent probe for detecting hypochlorite in naked eyes and in real time can keep a stable structure in different pH value ranges. Adding ClO – (5.0×10 –4 mol/L), clO is carried out at a pH in the range of 4 to 12 – The fluorescence of the fluorescent probe solution can be reduced by 4.0 times. That is, the change in pH value recognizes ClO with respect to the fluorescence intensity of the fluorescent probe solution – Fluorescent intensity of (C)No influence is generated on ClO of the fluorescent probe – Can be realized within a pH range of 4 to 12.
Fluorescent probe solution for naked eye and real-time detection of hypochlorite prepared in example 1 (10. Mu. Mol.L –1 ) Detection of ClO – (10μmol·L –1 ) The change with time of the fluorescence intensity of (c) is shown in FIG. 6. The synthesized hypochlorite fluorescent probe changes rapidly from pink to green after reacting with hypochlorite, and exhibits obvious fluorescence quenching.
Based on the above results, in ClO – The concentration is 0 to 1 multiplied by 10 –6 In the mol/L range, the fluorescent probe for naked eye and real-time detection of hypochlorite prepared in example 1 was prepared at 615nm (lambda ex =420 nm) fluorescence emission intensity and ClO – The standard curve of the concentration change is shown in fig. 7. According to the calculation formula of the detection limit, a blank parallel experiment is adopted to obtain a linear regression equation y= -13.54x+801.78 through fitting, and the standard deviation R is calculated 2 = 0.9940 naked eye and real-time detection of hypochlorite fluorescent probe pair ClO – The detection limit of (2) reaches 46.19nmol/L. The embodiment is illustrated that the preparation of naked eyes and the real-time detection of hypochlorite fluorescent probes can realize the ClO of the aqueous solution – Is a quantitative detection of (a).
Example 2: the preparation method of the fluorescent probe for detecting hypochlorite in real time and with naked eyes in the embodiment comprises the following steps:
1. weighing 0.32g of 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde and 0.20mL of 4-methylacetophenone, adding into a three-mouth bottle with a reflux device, adding 50mL of ethanol as a reaction solvent, heating to boiling and refluxing under stirring, and keeping for 6 hours for reaction; cooling to room temperature after the reaction is finished to obtain yellow liquid;
2. adding distilled water into the yellow liquid, regulating the pH value to be neutral by acetic acid, adding NaCl solution with the mass percent concentration of 20%, standing, and separating out solids; filtering out the solid, and drying to obtain a crude product;
3. the crude product was recrystallized from ethyl acetate to give a fluorescent probe for naked eye and real-time detection of hypochlorite, which was a dark yellow solid.
The yield of the fluorescent probe for detecting hypochlorite in real time with naked eyes prepared in the embodiment is 85%.
Example 3: the preparation method of the fluorescent probe for detecting hypochlorite in real time and with naked eyes in the embodiment comprises the following steps:
1. weighing 0.32g of 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde and 0.33mL of 4-methylacetophenone, adding into a three-necked flask with a reflux device, adding 40mL of ethyl acetate as a reaction solvent, heating to boiling and refluxing under stirring, and keeping for 3 hours for reaction; cooling to room temperature after the reaction is finished to obtain yellow liquid;
2. adding distilled water into the yellow liquid, regulating the pH value to be neutral by acetic acid, adding NaCl solution with the mass percent concentration of 20%, standing, and separating out solids; filtering out the solid, and drying to obtain a crude product;
3. the crude product was recrystallized from methanol to give a fluorescent probe for naked eye and real-time detection of hypochlorite, which was a dark yellow solid.
The yield of the fluorescent probe for detecting hypochlorite in real time with naked eyes prepared in the embodiment is 75%.
Example 4: the preparation method of the fluorescent probe for naked eye and real-time hypochlorite detection of the embodiment comprises the following steps of
1. Weighing 0.32g of 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde and 0.39mL of 4-methylacetophenone, adding into a three-necked flask with a reflux device, adding 50mL of dichloromethane as a reaction solvent, heating to boiling and refluxing under stirring, and keeping for 5 hours for reaction; cooling to room temperature after the reaction is finished to obtain yellow liquid;
2. adding distilled water into the yellow liquid, regulating the pH value to be neutral by acetic acid, adding NaCl solution with the mass percent concentration of 20%, standing, and separating out solids; filtering out the solid, and drying to obtain a crude product;
3. the crude product was recrystallized from ethanol to give a fluorescent probe for naked eye and real-time detection of hypochlorite, which was a dark yellow solid.
The yield of the fluorescent probe for detecting hypochlorite in real time with naked eyes prepared in the embodiment is 72%.
Example 5: the preparation method of the fluorescent probe for detecting hypochlorite in real time and with naked eyes in the embodiment comprises the following steps:
1. weighing 0.32g of 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde and 0.53mL of 4-methylacetophenone, adding into a three-mouth bottle with a reflux device, adding 60mL of acetone as a reaction solvent, heating to boiling reflux under stirring condition, and keeping for 8 hours for reaction; cooling to room temperature after the reaction is finished to obtain yellow liquid;
2. adding distilled water into the yellow liquid, regulating the pH value to be neutral by acetic acid, adding NaCl solution with the mass percent concentration of 20%, standing, and separating out solids; filtering out the solid, and drying to obtain a crude product;
3. the crude product was recrystallized from ethyl acetate to give a fluorescent probe for naked eye and real-time detection of hypochlorite, which was a dark yellow solid.
The yield of the fluorescent probe for detecting hypochlorite in real time with naked eyes prepared in the embodiment is 80%.
Example 6: the preparation method of the fluorescent probe for naked eye and real-time detection of hypochlorite in the embodiment comprises the following steps:
1. weighing 0.32g of 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde and 0.60mL of 4-methylacetophenone, adding into a three-necked flask with a reflux device, adding 60mL of acetonitrile as a reaction solvent, heating to boiling and refluxing under stirring, and keeping for 9 hours for reaction; cooling to room temperature after the reaction is finished to obtain yellow liquid;
2. adding distilled water into the yellow liquid, regulating the pH value to be neutral by acetic acid, adding NaCl solution with the mass percent concentration of 20%, standing, and separating out solids; filtering out the solid, and drying to obtain a crude product;
3. the crude product was recrystallized from propanol to give a fluorescent probe for naked eye and real-time detection of hypochlorite, which was a dark yellow solid.
The yield of the fluorescent probe for detecting hypochlorite in real time with naked eyes prepared in the embodiment is 70%.
Claims (9)
1. The fluorescent probe for detecting hypochlorite in real time with naked eyes is characterized by comprising the following structural formula:
2. a method for preparing a fluorescent probe for naked eye and real-time detection of hypochlorite in accordance with claim 1, characterized in that the method comprises the following steps:
1. the ratio of the amounts of the substances of 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde to 4-methylacetophenone was 1: (1-6) respectively weighing 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde v and 4-methylacetophenone, adding the mixture into a three-mouth bottle with a reflux device, adding an organic solvent I as a reaction solvent, heating to boiling and refluxing under stirring, keeping for 1-10 hours for reaction, cooling to room temperature after the reaction is finished, filtering to remove solid byproducts, collecting filtrate, adding distilled water into the filtrate, regulating the pH value to be neutral by using a 5% NaOH solution, precipitating solids, filtering, collecting filter cakes, and obtaining a crude product;
2. recrystallizing the crude product by using an organic solvent II to obtain a fluorescent probe for detecting hypochlorite in real time with naked eyes, wherein the fluorescent probe is a dark yellow solid.
3. The method for preparing fluorescent probes for naked eye and real time detection of hypochlorite according to claim 2, wherein the organic solvent I in the first step is methanol, ethanol, ethyl acetate, acetonitrile, dichloromethane, glacial acetic acid or acetone.
4. The method for preparing fluorescent probe for naked eye and real time detection of hypochlorite according to claim 2 or 3, wherein the organic solvent II in the third step is ethanol, methanol, propanol, chloroform, dichloromethane or ethyl acetate.
5. The method for preparing a fluorescent probe for naked eye and real-time detection of hypochlorite according to claim 2 or 3, wherein the preparation method of 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde in the first step is as follows:
(1) The ratio of the amounts of the substances is 1: (1-3): (1-5) weighing phenanthrenequinone, terephthalaldehyde and ammonium acetate, adding the phenanthrenequinone, the terephthalaldehyde and the ammonium acetate into an organic solvent A as a reaction solvent, stirring at room temperature for 5-20 hours, and stopping the reaction; wherein the organic solvent A is benzene, chlorobenzene, glacial acetic acid or N, N-dimethylformamide;
(2) Distilled water is added into the reaction system, the pH value of the reaction system is regulated to be neutral by using a NaOH solution with the concentration of 0.1mol/L, solids are separated out, suction filtration is carried out, and a filter cake is collected, so that a crude product is obtained;
(3) Adding the crude product into an organic solvent B, heating to the reflux temperature of the solvent, recrystallizing, filtering while the crude product is hot, and drying to obtain 4 (1-H-phenanthro [9,10-d ] imidazole-2) -benzaldehyde; wherein the organic solvent B is ethyl acetate, dichloromethane, chloroform, acetone, ethanol or methanol.
6. Use of a fluorescent probe for naked eye and real time detection of hypochlorite ions according to claim 1, characterized in that the use of the fluorescent probe is for qualitative or quantitative detection of hypochlorite ions in a solution.
7. The use of a fluorescent probe for naked eye and real time detection of hypochlorite according to claim 6, wherein the fluorescent probe for naked eye and real time detection of hypochlorite is used for ClO in aqueous solution – The naked eye qualitative detection method comprises the following steps:
1. dissolving a fluorescent probe for detecting hypochlorite in real time with naked eyes in an organic solvent to prepare a probe stock solution A;
2. diluting the probe stock solution A obtained in the step one to a concentration of 0.01-0.05 mmol.L by using PBS phosphate buffer solution –1 Probe with a probe tipSolution B;
3. adding a solution to be tested into the probe solution B in the second step to obtain a test solution C;
4. after the test solution C stands for 2-3 min, observing the color of the test solution C under natural light, and judging that the solution to be tested contains ClO if the color is changed from yellow to colorless – The method comprises the steps of carrying out a first treatment on the surface of the Or after the test solution C stands for 2-3 min, observing the color of the test solution C under the irradiation of a handheld ultraviolet lamp, and judging that the solution to be tested contains ClO if the color is changed from pink to green – 。
8. The use of a fluorescent probe for naked eye and real time detection of hypochlorite in solution according to claim 6, wherein the use of the fluorescent probe for naked eye and real time detection of ClO in solution – The fluorescence qualitative detection method of (2) comprises the following steps:
1. dissolving a fluorescent probe for detecting hypochlorite in real time with naked eyes in an organic solvent to prepare a probe stock solution A;
2. diluting the probe stock solution A obtained in the step one to a concentration of 0.01-0.05 mmol.L by using PBS phosphate buffer solution –1 A probe solution B;
3. measuring fluorescence emission spectrum of probe solution B with 420nm as excitation wavelength, recording emission intensity at 615nm, and recording as T B ；
4. Adding a solution to be tested into the probe solution B in the second step to obtain a test solution C;
5. after the test solution C is kept stand for 2 to 3 minutes, the fluorescence emission spectrum of the test solution C is measured by taking 420nm as an excitation wavelength, and the emission intensity when the emission wavelength is 615nm is recorded and is recorded as T C ；
6. Comparison T B And T C If 4T B ≤T C Judging that the solution to be tested contains ClO – 。
9. The use of a fluorescent probe for naked eye and real time detection of hypochlorite according to claim 6, wherein the naked eye and real time detection is usedClO in hypochlorite fluorescent probe pair solution – The fluorescent quantitative detection method of (2) is a standard curve method, and is specifically carried out according to the following steps:
1. dissolving a fluorescent probe for detecting hypochlorite in real time with naked eyes in an organic solvent to prepare a probe stock solution A;
2. diluting the probe stock solution A obtained in the step one to a concentration of 0.01-0.05 mmol.L by using PBS phosphate buffer solution –1 A probe solution B;
3. gradually adding the probe solution B to the probe solution B at the concentration of C respectively 1 、C 2 、C 3 ……C n ClO of (C) – Solutions, respectively obtaining sample solutions B 1 、B 2 、B 3 ……B n ；
4. Measurement of sample solution B with excitation wavelength of 420nm 1 ～B n The fluorescence emission spectra of (2) were recorded as T, and the emission intensity at 615nm was recorded, respectively B1 ～T Bn ；
5. In ClO – Concentration C of (2) 1 ～C n On the abscissa, with the emission intensity T of the sample solution B1 ～T Bn Plotted as ordinate, clO – A standard curve between concentration-fluorescence intensity;
6. adding the solution to be tested into the probe solution B, and uniformly mixing to prepare a test solution B x ；
7. 2.0. Mu.M ClO was added to the test solution Bx – Measuring fluorescence emission spectrum of the test solution Bx by using 420nm as excitation wavelength, recording emission intensity at 615nm, and recording as T Bx ；
8. Will T Bx Looking up T by comparing the ordinate of the standard curve in the fifth step Bx ClO in standard curve of intensity correspondence – Determining ClO in the solution to be tested according to the recovery rate and average deviation after 3 times of measurement – Concentration.
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