CN117447392B - A fluorescent material and its preparation method and application - Google Patents
A fluorescent material and its preparation method and application Download PDFInfo
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- CN117447392B CN117447392B CN202311184633.8A CN202311184633A CN117447392B CN 117447392 B CN117447392 B CN 117447392B CN 202311184633 A CN202311184633 A CN 202311184633A CN 117447392 B CN117447392 B CN 117447392B
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- 239000000463 material Substances 0.000 title claims abstract description 40
- 238000002360 preparation method Methods 0.000 title claims abstract description 7
- 238000001514 detection method Methods 0.000 claims abstract description 27
- -1 uranyl ions Chemical class 0.000 claims abstract description 21
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 36
- 229940098773 bovine serum albumin Drugs 0.000 claims description 32
- 239000007850 fluorescent dye Substances 0.000 claims description 27
- 238000000034 method Methods 0.000 claims description 27
- 150000001875 compounds Chemical class 0.000 claims description 23
- 230000008569 process Effects 0.000 claims description 15
- NBIIXXVUZAFLBC-UHFFFAOYSA-L Phosphate ion(2-) Chemical compound OP([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-L 0.000 claims description 7
- JUIKUQOUMZUFQT-UHFFFAOYSA-N 2-bromoacetamide Chemical compound NC(=O)CBr JUIKUQOUMZUFQT-UHFFFAOYSA-N 0.000 claims description 4
- 238000012921 fluorescence analysis Methods 0.000 claims description 3
- LMBFAGIMSUYTBN-MPZNNTNKSA-N teixobactin Chemical compound C([C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@H](CCC(N)=O)C(=O)N[C@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@H]1C(N[C@@H](C)C(=O)N[C@@H](C[C@@H]2NC(=N)NC2)C(=O)N[C@H](C(=O)O[C@H]1C)[C@@H](C)CC)=O)NC)C1=CC=CC=C1 LMBFAGIMSUYTBN-MPZNNTNKSA-N 0.000 claims description 3
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- 125000003368 amide group Chemical group 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
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- AYEKOFBPNLCAJY-UHFFFAOYSA-O thiamine pyrophosphate Chemical compound CC1=C(CCOP(O)(=O)OP(O)(O)=O)SC=[N+]1CC1=CN=C(C)N=C1N AYEKOFBPNLCAJY-UHFFFAOYSA-O 0.000 description 10
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
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- 102000007562 Serum Albumin Human genes 0.000 description 4
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 4
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- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- FKNQCJSGGFJEIZ-UHFFFAOYSA-N 4-methylpyridine Chemical compound CC1=CC=NC=C1 FKNQCJSGGFJEIZ-UHFFFAOYSA-N 0.000 description 2
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 2
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- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 150000003869 acetamides Chemical class 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
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- 229940061607 dibasic sodium phosphate Drugs 0.000 description 1
- WYICGPHECJFCBA-UHFFFAOYSA-N dioxouranium(2+) Chemical compound O=[U+2]=O WYICGPHECJFCBA-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- CBMPTFJVXNIWHP-UHFFFAOYSA-L disodium;hydrogen phosphate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].OP([O-])([O-])=O.OC(=O)CC(O)(C(O)=O)CC(O)=O CBMPTFJVXNIWHP-UHFFFAOYSA-L 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- VANPZBANAIIRJW-UHFFFAOYSA-N diuranium Chemical compound [U]#[U] VANPZBANAIIRJW-UHFFFAOYSA-N 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000000695 excitation spectrum Methods 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 238000002795 fluorescence method Methods 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
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- 239000003068 molecular probe Substances 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 229910000510 noble metal Inorganic materials 0.000 description 1
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- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
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- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
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- 239000006223 plastic coating Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- JUJWROOIHBZHMG-UHFFFAOYSA-O pyridinium Chemical compound C1=CC=[NH+]C=C1 JUJWROOIHBZHMG-UHFFFAOYSA-O 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000006862 quantum yield reaction Methods 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000002285 radioactive effect Effects 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 238000003950 stripping voltammetry Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 1
- 125000005289 uranyl group Chemical group 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
- C07D213/36—Radicals substituted by singly-bound nitrogen atoms
- C07D213/38—Radicals substituted by singly-bound nitrogen atoms having only hydrogen or hydrocarbon radicals attached to the substituent nitrogen atom
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6432—Quenching
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Optics & Photonics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Materials Engineering (AREA)
- Engineering & Computer Science (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
本发明公开了一种荧光材料及其制备方法和应用，本发明以三苯胺TPA荧光基团为发光色团，引入具有与铀酰离子配位潜力的酰胺基团和可以增大探针水溶性的吡啶盐离子季铵盐，合成一种新型AIE荧光材料TPP‑BAC，不但原料廉价易得、反应简单、产率高、细胞毒性低，而且可以在纯水体系以及细胞中实现原位、高灵敏性、快速的铀检测。The invention discloses a fluorescent material and a preparation method and application thereof. The invention uses a triphenylamine TPA fluorescent group as a luminescent chromophore, introduces an amide group with a potential for coordination with uranyl ions and a pyridinium ion quaternary ammonium salt that can increase the water solubility of the probe, and synthesizes a novel AIE fluorescent material TPP-BAC. The novel AIE fluorescent material TPP-BAC has cheap and readily available raw materials, simple reactions, high yields, and low cytotoxicity, and can realize in-situ, highly sensitive, and rapid uranium detection in a pure water system and cells.
Description
Technical Field
The invention relates to the technical field of fluorescence detection, in particular to a fluorescent material and a preparation method and application thereof.
Background
Uranium (Uranium) is a natural radioactive element that plays an important role in nuclear fuels and weapons. The fuel currently used in nuclear power plants is mainly uranium due to its unique advantages. However, the development and utilization of such high-energy sources is accompanied by a great hazard to the environment, living things, etc., due to the radioactivity and chemical toxicity of uranium. Uranium exists in a variety of oxidation states (from +2 to +6), whereas uranyl ions (UO 2 2+) are the most stable chemical form of uranium in aerobic environmental conditions, and are the predominant form of uranium present under acidic conditions. Currently heap leaching and in situ leaching techniques are popular methods of recovering and extracting uranium. Acid is a common means of both of these techniques, but this also makes the surrounding environment acidic, making UO 2 2+ easily mobile in the environment. And when uranium enters the human body, it accumulates in human tissues and organs and generates internal radiation, causing irreversible damage to humans or other organisms, resulting in various diseases such as kidney disease and osteosarcoma.
Along with the development and utilization of nuclear energy, the rapid detection of uranyl ions in environmental water is important in consideration of environmental hazard and potential human health threat. The detection mode of uranium at present mainly utilizes large-scale instruments such as an inductively coupled plasma mass spectrometer and is limited to a field sampling-laboratory analysis mode, so that the requirements of field in-situ uranium detection, pollution early warning and emergency detection which are continuously improved are difficult to meet. Therefore, various novel sensitive in-situ or remote detection methods are developed, and the method has important significance for the real-time monitoring of uranium and other nuclide pollution and the effective utilization of nuclear resources.
Traditional uranium detection methods, such as ICP-MS, ICP-AES, solid fluorescence method, stripping voltammetry and the like, often depend on large precise instruments, and require complex pretreatment, high cost and complex operation. Fluorescence spectroscopy (Fluorescence Spectrometry) is an effective way to perform qualitative and quantitative analysis on various pollutants in environmental samples, and is rapidly becoming one of the mainstream uranyl ion detection methods due to higher sensitivity and selectivity. The basic principle is that an optical sensor is built by constructing a fluorescent probe capable of specifically interacting with a target analyte according to mechanisms such as light-induced electron transfer (Photoinduced electron transfer, PET), aggregation-induced emission (AIE), fluorescence resonance energy transfer (Fluorescence resonance ENERGY TRANSFER, FRET) and the like, the fluorescent probe is excited by using a beam of excitation light with a specific wavelength, and then corresponding fluorescent optical property changes (such as light intensity, wavelength shift and the like) are measured, and the fluorescent probe is converted into specific signals such as electric signals and the like through the sensor to detect. Compared with other detection methods, the fluorescence spectrometry does not need expensive instruments, has the characteristics of simple operation, high efficiency, high sensitivity, low cost, convenience and the like, and can even realize naked eye identification. This is also a prerequisite for the rapid development of various fluorescent materials at present. Aggregation-induced emission (AIE), a fluorescent phenomenon that is diametrically opposed to the properties of conventional organic fluorescent materials (Aggregation caused quenching, ACQ). Compared with the traditional organic fluorescent material, the AIE material has the remarkable characteristics of high quantum yield, large Stokes shift, almost no overlapping of excitation and emission spectra, low cytotoxicity and the like in a polymerized state. Currently, AIE fluorescent materials, which change fluorescence emission properties by generating strong chelation with metal ions and the like to realize sensing of target metals, have attracted much interest of scientific researchers and have been developed for sensors of various heavy metal pollutants. Some AIE fluorescent materials have been designed for UO 2 2+ detection, but most of the materials still cannot meet the purpose of field in-situ detection, and most of the AIE fluorescent materials are insoluble in water and require the addition of an organic solvent to realize, so that the AIE fluorescent materials are limited in practical application. Most AIE fluorescent probes need toxic organic solvents in practical application, so that the health of monitoring personnel is affected, the environment is polluted, and the existence of the organic solvents can damage cells and influence the intracellular experimental results.
Disclosure of Invention
The present invention aims to solve at least one of the above technical problems in the prior art. To this end, the invention aims to provide a fluorescent material, a preparation method and application thereof.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
in a first aspect of the invention, there is provided a compound of formula I:
in a second aspect of the present invention, there is provided a process for the preparation of the compound of formula I, comprising the steps of:
The compound of formula II And reacting with halogenated acetamide to obtain the compound of formula I.
In some embodiments of the invention, the molar ratio of the compound of formula II to haloacetamide is 1: (1-3).
In some embodiments of the invention, the solvent of the reaction comprises any one of ethyl acetate, tetrahydrofuran.
In some embodiments of the invention, the haloacetamide comprises a bromoacetamide such as 2-bromoacetamide.
In a third aspect of the invention, there is provided a fluorescent material comprising the compound of formula I.
In a fourth aspect of the invention, there is provided a fluorescent probe comprising a compound of formula I or the fluorescent material.
In some embodiments of the invention, the fluorescent probe further comprises bovine (and/or human) serum albumin and/or hydrogen phosphate ions. Preferably, the fluorescent probe is capable of producing a sensitive response to uranyl ions in a pure water system in the presence of bovine (and/or human) serum albumin and/or hydrogen phosphate ions.
In some embodiments of the invention, the hydrogen phosphate ion comprises H 2PO4 - and/or HPO 4 2-.
In some embodiments of the invention, the ratio of the compound of formula I to bovine (and/or human) serum albumin is 1. Mu. Mol (0.01-3) g; preferably 1. Mu. Mol (0.5 to 2.5) g.
In a fifth aspect of the invention, a fluorescent sensing system is provided comprising said fluorescent probe.
In some embodiments of the invention, the fluorescence sensing system comprises a fluorescence probe and a fluorescence spectrometer.
In some embodiments of the invention, the fluorescent probe is in series with the fluorescence spectrometer.
In some embodiments of the invention, the fluorescence sensing system comprises a fluorescence probe, a probe head, a laser light source, a Y-type optical fiber, a flange adapter and a fiber optic spectrometer.
In some embodiments of the present invention, in the fluorescence sensing system, the Y-type optical fiber and the probe are detachably connected through a flange adapter.
In some embodiments of the present invention, in the fluorescence sensing system, the laser light source, the fluorescent probe and the optical fiber spectrometer are sequentially connected through a Y-shaped optical fiber; wherein, the Y-shaped optical fiber is detachably connected with the probe through the flange adapter.
In some embodiments of the present invention, in the fluorescence sensing system, the laser emitted by the laser light source is transmitted to the fluorescent probe through the Y-shaped optical fiber, and the fluorescent signal of the fluorescent probe is reflected back to the Y-shaped optical fiber and transmitted to the optical fiber spectrometer through the Y-shaped optical fiber.
In some embodiments of the invention, the probe is a fiber optic probe.
In some embodiments of the invention, the fluorescence sensing system further comprises a computer, and the fiber optic spectrometer is connected to the computer.
In some embodiments of the invention, the fluorescence sensing system further comprises a filter disposed between the Y-fiber and the fiber optic spectrometer, the Y-fiber being connected to the fiber optic spectrometer by the filter.
In a sixth aspect of the invention there is provided a method of detecting uranyl ions comprising contacting uranyl ions with a system comprising said compound of formula I for fluorescence analysis.
In some embodiments of the invention, the analysis comprises fluorescence intensity analysis and/or fluorescence imaging analysis.
In some embodiments of the invention, the method of detecting uranyl ions comprises contacting uranyl ions with a system comprising the compound of formula I and/or bovine (and/or human) serum albumin and/or hydrogen phosphate ions for fluorescence intensity analysis.
In some embodiments of the invention, the hydrogen phosphate ion comprises H 2PO4 - and/or HPO 4 2-.
In some embodiments of the invention, the method of detecting uranyl ions comprises subjecting cells to a fluorescence imaging analysis after incubating the cells in a system comprising the compound of formula I and bovine serum albumin and a system comprising uranyl ions sequentially.
In some embodiments of the invention, the system further comprises a buffer solution; preferably, the pH of the buffer solution is 7 to 8.
In some embodiments of the invention, the system comprises a solution system and/or a cell system.
In some embodiments of the invention, the solution system comprises a water sample system, such as an environmental real water sample system.
In some embodiments of the invention, fluorescence analysis of a sample containing uranyl ions is included using the fluorescent probe or the fluorescent sensing system.
In a seventh aspect, the invention provides an application of the compound of formula I or the fluorescent material or the fluorescent probe or the fluorescent sensing system in detection of uranyl ions.
The beneficial effects of the invention are as follows:
1. the invention provides a method for realizing in-situ, high-sensitivity and rapid uranium detection.
2. The invention takes triphenylamine TPA fluorescent group as a luminescent group, introduces an amide group with coordination potential with uranyl ions and pyridinium ionic quaternary ammonium salt capable of increasing water solubility to synthesize the novel AIE fluorescent material TPP-BAC, and has the advantages of low cost and easy obtainment of raw materials, simple reaction, high yield and good biocompatibility.
3. The invention successfully establishes a novel portable fluorescent optical fiber sensing system PFFS (Portable Fluorescence Fiber Sensor), and can realize detection exploration of target analytes in the field in situ.
4. The invention uses bovine serum albumin BSA to induce aggregation of TPP-BAC, the TPP-BAC is a micromolecular AIE probe, the molecular size is much smaller than that of noble metal nano particles, the molecular probe TPP-BAC aggregates in a hydrophobic cavity of BSA molecules, the fluorescence intensity is increased due to limited movement, the high-sensitivity detection of uranyl ions in a complete aqueous solution can be realized, and the use of toxic and harmful organic solvents is avoided.
5. The invention can realize the high-sensitivity imaging detection exploration of trace uranium acyl ions in cells and carry out risk early warning on uranium pollution of human bodies.
Drawings
FIG. 1 is a schematic diagram showing the synthesis route of TPP-BAC (Compound of formula I) in example 1 of the present invention.
Fig. 2 is a high resolution mass spectrum of the fluorescent material TPP in example 1 of the present invention.
FIG. 3 is a nuclear magnetic resonance spectrum of the fluorescent material TPP in example 1 of the present invention.
FIG. 4 is a nuclear magnetic resonance spectrum of the fluorescent material TPP in example 1 of the present invention.
FIG. 5 is a high-resolution mass spectrum of a fluorescent material TPP-BAC in example 1 of the present invention.
FIG. 6 is a nuclear magnetic resonance hydrogen spectrum of a fluorescent material TPP-BAC in example 1 of the present invention.
FIG. 7 is a nuclear magnetic resonance spectrum of a fluorescent material TPP-BAC in example 1 of the present invention.
FIG. 8 is a graph of the ultraviolet absorption spectrum and the optimal fluorescence emission spectrum of TPP-BAC.
FIG. 9 is a graph of fluorescence spectra of TPP-BAC in various ethyl acetate/ethanol mixtures.
FIG. 10 shows the effect of different buffer solvents on TPP-BAC fluorescence intensity and its selectivity for detecting UO 2 2+.
FIG. 11 is a graph showing changes in fluorescence intensity of TPP-BAC after aggregation in bovine serum albumin.
FIG. 12 is a graph showing (a) fluorescence spectrum and (b) peak change of optimal emission at 575nm of TPP-BAC with change in BSA concentration.
FIG. 13 is a graph showing changes in fluorescence intensity of TPP-BAC solutions after various metal ions are added.
FIG. 14 is a graph showing the change in fluorescence response of TPP-BAC to UO 2 2+ under the interference of (a) metal ions and (b) anions.
FIG. 15 shows the cell viability of HeLa cells detected by CCK-8.
FIG. 16 is a graph showing changes in HeLa intracellular fluorescence after treatment with TPP-BAC and UO 2 2+ at different concentrations, respectively; channels i, ii, iii represent cell imaging plots of red, combined and bright field, respectively; a, B, C, D, E each represent a different concentration of UO 2 2+ added.
FIG. 17 is a graph of a linear fit between mean gray scale values of cells treated with TPP-BAC and UO 2 2+ concentration.
FIG. 18 emission spectra of (a) TPP-BAC@FL-2700 in the presence of various concentrations of UO 2 2+; (b) TPP-BAC@FL-2700 versus UO 2 2+ concentration graph (built-in graph is a linear fit curve).
FIG. 19 is a block diagram of a portable fluorescence fiber optic sensing system.
FIG. 20 is a physical construction diagram of a portable fluorescence fiber optic sensing system; (a) a PFFS physical whole shooting picture; (b) a Y-type fiber optic connection node; (c) an optical fiber probe and a self-built detection platform.
FIG. 21 is an emission spectrum of (a) TPP-BAC@PFFS in the presence of various concentrations of UO 2 2+; (b) TPP-BAC@PFFS fluorescence intensity versus UO 2 2+ concentration (built-in graph is a linear fit curve).
Detailed Description
The present invention will be described in further detail with reference to specific examples. The starting materials, reagents or apparatus used in the examples and comparative examples were either commercially available from conventional sources or may be obtained by prior art methods unless specifically indicated. Unless otherwise indicated, assays or testing methods are routine in the art.
Example 1
The compound of formula I is prepared in this example by the following steps:
FIG. 1 shows a schematic diagram of the synthetic route of TPP-BAC (Compound of formula I).
S1: synthesis of triphenylamine fluorescent molecule TPP: diphenylamino-4-benzaldehyde (4.8 mmol,1.3125 g), 4-methylpyridine (7.2 mmol,0.7 mL) were added to a round bottom flask containing a mixed solvent of acetic anhydride (30 mL)/acetic acid (10 mL) and stirred under reflux at 150℃under nitrogen for 12h. After the reaction was completed, after cooling to room temperature, the reaction solution was spin-dried at 80℃to remove acetic acid and acetic anhydride, and then the crude product was purified by column chromatography using petroleum ether/ethyl acetate (25:1, v/v) mixed eluent to give TPP as an orange-yellow solid in a yield of 51%.
S2 Synthesis of TPP-BAC (Compound of formula I): TPP (1 mmol,0.3482 g) obtained in S1 and 2-bromoacetamide (1.2 mmol,0.1643 g) were dissolved in a round bottom flask containing ethyl acetate (20 mL) and refluxed under nitrogen for 2h. After the reaction was completed and cooled to room temperature, the product was filtered to give a red solid, which was recrystallized using a petroleum ether/ethyl acetate (2:1, v/v) mixed solvent, and the obtained crystals were dried under vacuum in 87% yield.
The prepared TPP and TPP-BAC are characterized.
Fig. 2, 3 and 4 are respectively a high-resolution mass spectrum, a nuclear magnetic hydrogen spectrum and a nuclear magnetic carbon spectrum of the fluorescent material TPP.
Nuclear magnetic resonance hydrogen spectrum analysis of TPP ：1H NMR(600MHz,CDCl3):δ＝11.13(s,1H),9.89(s,1H),7.58(d,J＝8.1Hz,1H),7.54-7.47(m,2H),7.32-7.28(m,4H),7.23(dd,J＝8.1,1.6Hz,1H),7.18(d,J＝1.5Hz,1H),7.16-7.10(m,6H),7.10-7.05(m,2H)mg/L.
Nuclear magnetic resonance carbon spectrum analysis of TPP ：13C NMR(151MHz,CDCl3):δ＝195.76,162.04,148.82,147.24,134.08,132.14,129.44,128.07,125.04,123.62,122.75,119.18,118.20,114.78.
Fig. 5, 6 and 7 are respectively high-resolution mass spectrum, nuclear magnetic hydrogen spectrum and nuclear magnetic carbon spectrum of a fluorescent material TPP-BAC (compound of formula I).
Nuclear magnetic resonance hydrogen spectrometry of TPP-BAC (Compound of formula I) ：1H NMR(600MHz,DMSO):δ＝12.97(s,0H),8.87(s,0H),7.78-7.57(m,2H),7.38-7.33(m,2H),7.28(dd,J＝8.1,1.7Hz,1H),7.22(d,J＝1.6Hz,1H),7.15(dd,J＝7.9,1.1Hz,1H),7.13-7.07(m,3H),7.06-6.99(m,2H),6.79(dd,J＝8.0,1.2Hz,0H),6.73-6.49(m,0H),5.08(s,1H)mg/L.
Nuclear magnetic resonance carbon spectrum analysis of TPP-BAC (Compound of formula I) ：13C NMR(151MHz,DMSO):δ＝161.16-160.37(m,-1H),161.16-160.29(m,-1H),161.16-160.23(m,-1H),148.37-147.59(m,-1H),147.33(s,0H),144.33-143.83(m,0H),143.32-142.88(m,-1H),134.87-134.44(m,0H),133.49-132.41(m,-1H),130.15(s,1H),128.32(s,0H),124.97(s,1H),124.04(s,1H),123.10(s,0H),123.10(s,-5H),119.48-118.89(m,-1H),118.77(s,0H),117.33(d,J＝42.5Hz,0H),116.21-115.57(m,0H),116.21-115.44(m,0H),113.81(s,-1H).
Test example 1
The test example tests the fluorescence performance of the fluorescent material TPP-BAC, and comprises the following specific processes:
A certain amount of fluorescent material TPP-BAC is dissolved in deionized water to prepare a 10 mu M solution for standby. First, scanning of ultraviolet absorption spectrum is performed: taking a proper amount of 10 mu M TPP-BAC aqueous solution in a cuvette, taking deionized water as a reference, scanning an absorption spectrum in a wavelength range of 700 nm-250 nm, positioning a plurality of absorption peaks (lambda 1,λ2 … …) according to the spectrum, and determining the optimal excitation wavelength lambda EXmax. Then, a proper amount of bovine serum albumin BSA was taken in the prepared 10. Mu.M TPP-BAC solution to prepare 2.0g/L BSA, 10. Mu.M TPP-BAC aqueous solution. Subsequently, according to several absorption peaks (lambda 1,λ2 … …) and lambda max obtained by ultraviolet absorption spectrum, which are respectively set as Excitation wavelengths (specification, E x), fluorescence emission spectrum scanning is performed by using FL-2700, and the optimal fluorescence emission wavelength lambda EMmax is determined by comparison.
FIG. 8 is an ultraviolet absorption spectrum and an optimal fluorescence emission spectrum of TPP-BAC. The blue and purple curves are the optimal absorption spectrum (absorpance) and the optimal Emission spectrum (Emission) of 10 μ MTPP-BAC (0.01M PBS buffer, ph=7.4, containing 0.2g/L BSA), respectively, with fluorescence detection parameters: slit (width) 5nm, voltage 700V. From the results, it was found that the TPP-BAC had an optimal excitation wavelength of 470nm and an optimal emission wavelength of 575nm, and that the solution showed strong orange-red fluorescence.
Ethanol and ethyl acetate were selected as benign solvent and poor solvent, respectively, and the fluorescence change of TPP-BAC in different solvents was observed by adjusting the ratio of benign solvent (ethanol) and poor solvent (ethyl acetate). That is, a series of 10. Mu.M TPP-BAC solutions were prepared with a mixture of benign solvent (ethanol) and poor solvent (ethyl acetate) in varying proportions, wherein the poor solvent volume fraction (f EA) was between 0% and 99%.
FIG. 9 is a fluorescence spectrum of TPP-BAC when the volume fraction content (f EA) of ethyl acetate in ethyl acetate/ethanol mixed solvent is changed from 0% -99%. As shown in fig. 9, TPP-BAC showed very weak fluorescence intensity in pure ethanol, while with the addition of the poor solvent ethyl acetate, the fluorescence intensity started to increase, and when the ethyl acetate volume fraction exceeded 70%, the fluorescence intensity increased more significantly and peaked at fEA =95%. The results indicate that TPP-BAC has typical AIE properties and can fluoresce strongly orange-red when it aggregates in solution. It was confirmed that TPP-BAC exhibited a typical aggregation-induced emission AIE effect. And when the volume fraction of the poor solvent in the mixed solvent is continuously increased to 99%, the fluorescence intensity is slightly reduced.
Test example 2
The experimental example explores the influence of different buffer solvents (pH=7.4) on the fluorescence intensity of TPP-BAC and the selectivity detection UO 2 2 +, and the specific process is as follows:
As shown in FIG. 10, condition ① is the fluorescent behavior of TPP-BAC in different buffer solvents (F/F 0). After adding 1 time of TPP-BAC equivalent UO 2 2+ (condition ②) to two buffer solvent systems containing no H 2PO4 -/HPO4 2-, tris-HCl (Tris (hydroxymethyl) aminomethane-hydrochloric acid) and HEPES-NaOH (4-hydroxyethyl piperazine ethanesulfonic acid-sodium hydroxide), The fluorescence intensity of the fluorescent probe TPP-BAC does not appear quenching, And when 1-fold TPP-BAC equivalent UO 2 2+ and 50-fold equivalent Na 2HPO4 are added simultaneously (condition ③), The fluorescence intensity of the fluorescent probe TPP-BAC appears to be obviously quenched. The ionization distribution of the combined phosphate in aqueous solutions of different pH values shows that when the pH value of the solution is 7.4, the main components in the solution are HPO 4 2- and H 2PO4 -, which shows that when the novel AIE fluorescent probe TPP-BAC exists simultaneously with UO 2 2+ and H 2PO4 -/HPO4 2-, resulting in a significant fluorescence quenching response. In contrast, in both CA-DSP (CITRIC ACID-Dibasic Sodium Phosphate, disodium citrate-hydrogen phosphate) and PBS (Phosphate buffer solution) buffer solvent systems containing sufficient H 2PO4 -/HPO4 2-, after 1-fold TPP-BAC equivalent of UO 2 2+ (condition ②) was added, The fluorescence intensity of the fluorescent probe TPP-BAC has obvious quenching effect, and the quenching rate is obviously higher than that of the former two buffer solvent systems without H 2PO4 -/HPO4 2-; After adding 1 time of TPP-BAC equivalent of UO 2 2+ and 50 times of Na 2HPO4 simultaneously to CA-DSP and PBS (condition ③), The fluorescence intensity of the fluorescent probe TPP-BAC also showed a stronger quenching effect, but at this time the quenching rate of condition ③ was lower than that of condition ②, probably because after an additional 50-fold equivalent of Na 2HPO4 was added to CA-DSP and PBS, Exceeding the buffer solvent buffer range (which is usually pka±1) results in a solution pH slightly above 7.4, and when the pH is above 7.4 the fluorescence intensity of TPP-BAC itself will increase slightly, resulting in a lower quenching rate.
Test example 3
The test example searches the fluorescence behavior of the fluorescent material TPP-BAC and searches whether Bovine Serum Albumin (BSA) has fluorescence enhancement effect on the TPP-BAC or not, and the specific process is as follows:
A quantity of bovine serum albumin was weighed and dissolved in PBS buffer (ph= 7.4,0.01M) to prepare a20 g/L BSA-PBS stock solution which was stored in a brown jar. A certain amount of TPP-BAC was then weighed and dissolved in water to prepare a stock solution of 100. Mu.M. Appropriate amounts of TPP-BAC stock solution and BSA-PBS stock solution were taken in a 5mL centrifuge tube, the volume was fixed to 4mL with PBS buffer (pH=7.4), and a TPP-BAC solution (10. Mu.M) with a BSA concentration of 1g/L was prepared while preparing a blank without adding BSA-PBS stock solution. The novel fluorescent probe TPP-BAC solution itself and the change of fluorescence intensity after BSA addition were investigated.
FIG. 11 shows the change in fluorescence intensity of TPP-BAC after aggregation in bovine serum albumin. It can be seen that the fluorescence intensity of TPP-BAC was very weak in pure PBS buffer solvent (pH=7.4), whereas with the addition of bovine serum albumin BSA, the fluorescence intensity of TPP-BAC was significantly enhanced, giving off strong orange-red fluorescence.
Test example 4
The test example explores the fluorescence behavior of the fluorescent material TPP-BAC, and explores the influence of Bovine Serum Albumin (BSA) with different concentrations on the fluorescence behavior of the TPP-BAC, and the specific process is as follows:
and (3) respectively placing a proper amount of TPP-BAC stock solution and BSA-PBS stock solution into a plurality of 5mL centrifuge tubes, fixing the volume to 4mL by using PBS buffer solution (pH value=7.4), respectively preparing TPP-BAC solution (10 mu M) with the BSA concentration of 0g/L、0.05g/L、0.1g/L、0.2g/L、0.4g/L、0.6g/L、0.8g/L、1.0g/L、1.2g/L、1.4g/L、1.6g/L、1.8g/L、2.0g/L、2.2g/L、2.5g/L、2.8g/L、3.0g/L, using 470nm as excitation wavelength and 575nm as emission wavelength, and using a FL-2700 fluorescence spectrometer to scan the fluorescence spectrum of the TPP-BAC solution.
FIG. 12 is a graph of (a) fluorescence spectrum of TPP-BAC (10. Mu.M) and (b) peak change of optimal emission at 575nm at a variation of bovine serum albumin concentration from 0 to 3.0 g/L. As can be seen, when the BSA concentration is 0-0.5 g/L, the fluorescence intensity of TPP-BAC is rapidly enhanced along with the continuous increase of the BSA concentration; when the BSA concentration reaches 0.5g/L, the TPP-BAC fluorescence amplification is gradually slowed down; at a BSA concentration of 2.0g/L, the TPP-BAC fluorescence reached a peak, at which time the TPP-BAC fluorescence intensity was enhanced by about 37.5 times as compared with that without BSA addition. Therefore, the BSA concentration in the TPP-BAC solution was set to 2.0g/L in other experiments.
Test example 5
The test example tests the selective response of the fluorescent material TPP-BAC to UO 2 2+, and the specific process is as follows:
Taking proper amounts of TPP-BAC stock solution and BSA-PBS stock solution in a plurality of 5mL centrifuge tubes, adding 1 time of TPP-BAC equivalent of different metal ions (UO2 2+、Cr3+、Be2+、Sr2+、Th4+、Cd2+、Cu2+、Na+、Mg2+、Ba2+、Ni2 +、Fe3+、Co2+、K+、Ca2+、Ag+、Hg2+), into the centrifuge tubes respectively, and using PBS buffer solution (pH value=7.4) to fix the volume to 4mL to prepare a mixed solution containing 1 time of TPP-BAC equivalent of different metal ions, 0.2g/L BSA and 10 mu M TPP-BAC, and simultaneously preparing a sample without adding any metal ions as a blank. After thoroughly mixing, the fluorescence emission spectrum of the solution was measured using FL-2700.
FIG. 13 is a graph showing changes in fluorescence intensity of TPP-BAC solutions after addition of various metal ions (10. Mu.M). It can be seen that other 16 metal ions have no significant fluorescence quenching effect on TPP-BAC, except UO 2 2+, indicating that the novel fluorescent probe TPP-BAC can achieve a selective response to UO 2 2+ in PBS buffer (0.01 m, ph=7.4).
A series of TPP-BAC solutions (10 μm, containing 2g/L BSA, PBS buffer, ph=7.4) containing 10 times each metal interfering ion (Cr3+,Be2+,Sr2+,Th4+,Na+,Mg2+,Ba2+,Ni2+,Fe3+,K+,Ca2+,Ag+,Hg2+) and 50 times TPA-SP equivalent of different anions (CH3COO-,NO3 -,NO2 -,SO4 2-,ClO4 -,F-,I-,Cl-) were prepared.
FIG. 14 shows the change in fluorescence response of TPP-BAC (10. Mu.M with 0.2g/L BSA, PBS buffer, pH=7.4) to UO 2 2+ (10. Mu.M) under interference of (a) metal ions (M n+, 100. Mu.M) and (b) anions (500. Mu.M). It can be seen that when 10-fold each of the metal interfering ions (Cr3+,Be2+,Sr2+,Th4+,Na+,Mg2+,Ba2+,Ni2+,Fe3+,K+,Ca2+,Ag+,Hg2+) and 50-fold each of the TPA-SP equivalent anions (CH3COO-,NO3 -,NO2 -,SO4 2-,ClO4 -,F-,I-,Cl-) were added to a10 μm TPP-BAC solution (0.2 g/L BSA,0.01M PBS buffer, ph=7.4), respectively, no significant interference was caused, indicating that the reaction between TPA-SP and UO 2 2+ had good anti-interference ability and was not interfered with by other possible coexisting ions.
Test example 6
The test example tests the cytotoxicity of the fluorescent material TPP-BAC, and the specific process is as follows:
The biocompatibility and cytotoxicity of TPP-BAC were determined by CCK-8 method. Cells were first digested and counted, and then a cell suspension was formulated at a concentration of 5X 10 4 cells/mL. mu.L of cell suspension (5X 10 3 cells per well) was added to each well of a 96-well cell culture plate, and the cell culture plate was placed in a 5% CO 2 incubator at 37℃overnight. Working solutions containing TPP-BAC reagents with different concentrations are respectively prepared by using a complete culture solution (90% DMEM culture solution and 10% fetal calf serum), 100 mu L of corresponding working solution is added into each hole after uniform mixing, so that the concentrations of the TPP-BAC are respectively 0, 1 mu M, 5 mu M, 10 mu M, 25 mu M and 50 mu M, and 3 concentrations are respectively arranged in parallel. After cells were cultured in a 5% CO 2 incubator at 37 ℃ for 48h, the supernatant was discarded, and then CCK-8 staining was performed on 96-well plates, i.e., 110 μl of CCK-8 working solution (V Culture solution ：V CCK-8 Stock solution =10:1) was added to each well, and after 2h of continuous culture in the incubator, the OD value (Optical density, λ=450 nm) of each well was read out using an enzyme-labeling instrument, and cell viability was calculated according to the following formula:
as a result, as shown in FIG. 15, there was no significant difference in cell proliferation after culturing at 37℃for 48 hours by adding TPP-BAC and a mixture of TPP-BAC and UO 2 2+ (molar ratio, 1:1) at different concentrations in the range of 0 to 50. Mu.M, respectively. Cell viability was slightly less than 90% at higher concentrations [ TPP-BAC+UO 2 2+ ] (50. Mu.M), with cell viability exceeding 91.22% at other conditions. The result shows that TPP-BAC has very low cytotoxicity to HeLa cells and can be used for subsequent imaging research of trace UO 2 2+ in cells.
Example 2
In the embodiment, the trace uranyl ions are detected in the cells by using a fluorescent material TPP-BAC, and the specific process is as follows:
In a cell imaging experiment, 10. Mu.M TPP-BAC solution (containing 0.2g/L BSA) was first prepared using PBS buffer (0.01M, pH=7.4). The treated HeLa cells were then seeded into confocal dishes at 37℃and incubated in 10. Mu.M TPP-BAC solution for 2h. Subsequently, after washing 3 times with PBS buffer (0.01M, ph=7.4) to remove excess TPP-BAC and other substances, incubation was continued for 2 hours with UO 2 2+ aqueous solutions (0 μm,1 μm,2 μm,4 μm,6 μm) of different concentrations, and washing 3 times again with PBS buffer (0.01M, ph=7.4). Immediately after the treatment, the cells were observed using a confocal laser microscope (Leica, SP 8) and the uptake of the sample by the cells was photographed. In the confocal shooting process, the wavelength of TPP-BAC emission light is 570 nm-580 nm (orange red).
FIG. 16 is a graph showing changes in HeLa intracellular fluorescence after treatment with TPP-BAC (10. Mu.M) and UO 2 2+ at different concentrations, respectively. TPP-BAC showed bright orange-red fluorescence in HeLa cells, channels i, ii, iii represent cell imaging patterns of red field (recording image signals of red wavelength 560 nm-680 nm), combined field (red field combined with bright field) and bright field (recording signals under natural light), respectively, and concentrations A, B, C, D, E represent added UO 2 2+ concentrations of 0. Mu.M, 1. Mu.M, 2. Mu.M, 4. Mu.M, 6. Mu.M, respectively. It was observed that with increasing concentrations of UO 2 2+ added to the cells, the orange-red fluorescence of TPP-BAC was gradually quenched and substantially quenched at a UO 2 2+ of 50. Mu.M.
FIG. 17 is a linear fit curve between the mean gray scale value (mean fluorescence intensity) and UO 2 2+ concentration of cells treated with TPP-BAC. The results of the cell experiments are highly consistent with those obtained by in vitro experiments. This shows that the probe has great detection potential for UO 2 2+ in vivo.
Example 3
The embodiment prepares a sensing system and tests the sensitivity thereof, and the specific process is as follows:
And (3) connecting a fluorescent probe TPP-BAC with a Hitachi (Hitachi) FL-2700 fluorescent spectrometer in series to obtain a TPP-BAC@FL-2700 sensing system.
In tandem with FL-2700, a series of 10. Mu.M TPP-BAC solutions (PBS buffer, pH=7.4) containing 0.2g/L BSA and 1000. Mu. M S 2O3 2- were prepared, followed by addition of stock solutions of UO 2 2+ at different concentrations, such that the concentrations of UO 2 2+ in the mixed solutions were 0.05. Mu.M, 0.1. Mu.M, 0.2. Mu.M, 0.4. Mu.M, 0.6. Mu.M, 0.8. Mu.M, 1. Mu.M, 2. Mu.M, 3. Mu.M, 4. Mu.M, 5. Mu.M, 6. Mu.M, 8. Mu.M and 10. Mu.M, respectively. Simultaneously, blank samples to be tested without adding UO 2 2+ are prepared, and each group of samples is parallel for 10 times. The fluorescence spectrum of the prepared sample was measured by using a Hitachi FL-2700 fluorescence spectrometer after shaking and homogenizing, and a linear relation curve of the fluorescence emission intensity of TPP-BAC with the concentration change of UO 2 2+ was drawn, and the result is shown in FIG. 18.
As can be seen from FIG. 18, the detection limit of the sensor on UO 2 2+ is 20.13nM, and the sensitivity is high.
Example 4
The embodiment prepares an optical fiber sensing system and tests the sensitivity thereof, and the specific process is as follows:
Fig. 19 and 20 are a structural view and a physical construction view of the portable fluorescent optical fiber sensor, respectively. The real object diagram can clearly show that the instrument has complete parts, simple construction and small occupied area, and can realize in-situ fluorescence detection by matching with the portable palm computer.
Construction of a TPP-BAC@PFFS sensing system obtained by connecting a portable fluorescent optical fiber sensor PFFS (portable fluorescent fiber sensor) with an AIE fluorescent probe TPP-BAC in series: the instrument structure of the portable fluorescent optical fiber sensor PFFS is formed by mutually assembling a laser light source, a Y-shaped optical fiber, a flange adapter, an optical filter, an optical fiber spectrometer, a long plastic cladding optical fiber, a computer and a fluorescent material TPP-BAC serving as a sensing unit. The components were assembled using SMA905 joints and a flange adapter was used to connect the Y-fiber with the fiber optic sensing probe. The fiber probe with the plastic coating removed is inserted into a solution containing AIE fluorescent material TPP-BAC for detection. The optical filter is arranged between the Y-shaped optical fiber and the spectrometer and is supported by the fixed support, so that light source peaks mixed in fluorescent signals are effectively filtered, and interference is eliminated. In order to explore the stability of the instrument and the feasibility of fluorescence detection, an optical fiber probe is inserted into TPA-P1 powder in an experiment, after the probe is static, fluorescence data is recorded once every 2s, the fluorescence data is recorded for 3 minutes, and the floating range of a fluorescence signal is observed and fitted.
In tandem with PFFS, a series of 10 μm TPP-BAC solutions (PBS buffer, ph=7.4) containing 0.2g/L BSA and 1000 μm M S 2O3 2- were prepared, followed by separate additions of stock solutions of UO 2 2+ at different concentrations, such that the concentration of UO 2 2+ in the mixed solution was 0.25 μm,0.5 μm,0.75 μm,1 μm,2 μm,3 μm,4 μm,5 μm,6 μm,8 μm and 10 μm, respectively. Simultaneously, blank samples to be tested without adding UO 2 2+ are prepared, and each group of samples is parallel for 10 times. The prepared sample is oscillated and shaken uniformly, then the fluorescence spectrum of the sample is measured by using a novel portable fluorescence optical fiber sensor (PFFS), and a linear relation curve of the fluorescence emission intensity of TPP-BAC along with the concentration change of UO 2 2+ is drawn, and the result is shown in figure 21.
As can be seen from FIG. 21, the detection limit of the sensor on UO 2 2+ is 134.36nM, and the sensitivity is high.
Example 5
In the embodiment, a TPP-BAC@FL-2700 sensing system obtained by connecting a TPP-BAC and a FL-2700 fluorescence spectrometer in series is adopted to detect an actual water sample, and the specific process is as follows:
while plotting the linear relationship, a 10. Mu.M TPP-BAC solution (PBS buffer, pH=7.4) containing 0.2g/L BSA and 1000. Mu. M S 2O3 2- was prepared, and 100. Mu.L of the treated actual water sample was added and the volume was fixed to 4mL with PBS buffer (pH=7.4). 3 replicates were prepared for each actual sample. The prepared sample was shaken and homogenized, and then its fluorescence spectrum was measured using a Hitachi (Hitachi) FL-2700 fluorescence spectrometer, and the average value of the measured fluorescence intensities was substituted into a linear relationship curve to calculate the concentration of UO 2 2+.
The results are shown in Table 1:
TABLE 1 determination of UO 2 2+ concentration in permeate and seawater samples Using TPP-BAC@FL-2700
As can be seen, the calculated UO 2 2+ concentrations for the TPP-BAC@FL-2700 detection permeate and seawater were 0.17 μM and 0.02 μM, respectively, and the sensor standard recovery rate and RSD obtained by the series connection of the TPP-BAC and FL-2700 fluorescence spectrometers were both within acceptable ranges.
Example 6
In the embodiment, a TPP-BAC@PFFS sensing system is adopted to detect an actual water sample, and the specific process is as follows:
While plotting the linear relationship, a 10. Mu.M TPP-BAC solution (PBS buffer, pH=7.4) containing 0.2g/L BSA and 1000. Mu. M S 2O3 2- was prepared, and 100. Mu.L of the treated actual water sample was added and the volume was fixed to 4mL with PBS buffer (pH=7.4). 3 replicates were prepared for each actual sample. And (3) after shaking the prepared sample, measuring the fluorescence spectrum of the sample by using a novel portable fluorescence optical fiber sensing system (PFFS), and substituting the average value of the measured fluorescence intensity into a linear relation curve to calculate the concentration of UO 2 2+.
The results are shown in Table 2:
TABLE 2 determination of UO 2 2+ concentration in permeate and seawater samples Using TPP-BAC@PFFS
As can be seen, the UO 2 2+ concentration calculated by the TPP-BAC@PFFS detection percolate and seawater is 0.17 mu M and undetected, and the sensor standard recovery rate and RSD obtained by the serial connection mode of the TPP-BAC and the PFFS portable fluorescent optical fiber sensor are both in an acceptable range.
Comparative example
The comparative example uses ICP-MS to detect the UO 2 2+ concentration of percolate and seawater, and the specific process is as follows:
A proper amount of uranium standard solution is taken to prepare a series of UO 2 2+ solutions with the concentration of 0.025 mu M, 0.05 mu M, 0.075 mu M, 0.1 mu M, 0.125 mu M, 0.15 mu M, 0.175 mu M and 0.2 mu M, and blank samples to be tested without adding UO 2 2+ are prepared. Each set of samples was run in parallel 3 times. And adjusting the ICP-MS instrument to an optimal working state, respectively measuring a standard solution and a blank solution, and drawing a linear relation curve. And then, carrying out digestion (V Sample of ：V Concentrated nitric acid = 10mL:1 mL) on the actual sample, carrying out constant volume to 10mL after digestion for 2h at 120 ℃, and measuring the actual sample by adopting an internal standard correction quantitative analysis method after cooling.
UO 2 2+ concentrations of the permeate and seawater were measured using ICP-MS and were 0.17. Mu.M and 0.02. Mu.M, respectively.
In conclusion, the concentration results of UO 2 2+ of the percolate and the seawater detected by using TPP-BAC@FL-2700, TPP-BAC@PFFS and ICP-MS are highly consistent, and the addition recovery rate and RSD of the sensors obtained by the two serial connection modes are within an acceptable range, so that the TPP-BAC@FL-2700 and TPP-BAC@PFFS sensing system built by the invention has great application potential in UO 2 2+ concentration detection of an actual water sample.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (7)
1. A compound of formula I:
。
2. a process for the preparation of a compound of formula I according to claim 1, wherein: the method comprises the following steps:
The compound of formula II And reacting with 2-bromoacetamide to obtain the compound of the formula I.
3. A fluorescent material characterized in that: comprising a compound of formula I as defined in claim 1.
4. A fluorescent probe, characterized in that: a fluorescent material according to claim 3.
5. The fluorescent probe of claim 4, wherein: the fluorescent probe also comprises bovine serum albumin and/or human serum albumin and/or hydrogen phosphate ions.
6. A detection method of uranyl ions is characterized in that: comprising contacting uranyl ions with a system comprising a compound of formula I as defined in claim 1 and hydrogen phosphate ions in vitro for fluorescence analysis.
7. Use of a compound of formula I according to claim 1 or a fluorescent material according to claim 3 or a fluorescent probe according to claim 4 for in vitro detection of uranyl ions.
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