WO2019218594A1 - Water-soluble phosphorescent nanoparticle for detecting hypochlorous acid using ratio method and preparation method and application thereof - Google Patents
Water-soluble phosphorescent nanoparticle for detecting hypochlorous acid using ratio method and preparation method and application thereof Download PDFInfo
- Publication number
- WO2019218594A1 WO2019218594A1 PCT/CN2018/111540 CN2018111540W WO2019218594A1 WO 2019218594 A1 WO2019218594 A1 WO 2019218594A1 CN 2018111540 W CN2018111540 W CN 2018111540W WO 2019218594 A1 WO2019218594 A1 WO 2019218594A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- hypochlorous acid
- water
- nanoparticle
- soluble
- complex
- Prior art date
Links
- 239000002105 nanoparticle Substances 0.000 title claims abstract description 53
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims abstract description 21
- 238000002360 preparation method Methods 0.000 title description 10
- 239000002202 Polyethylene glycol Substances 0.000 claims abstract description 4
- 229920001223 polyethylene glycol Polymers 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 18
- 150000001875 compounds Chemical class 0.000 claims description 13
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 12
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 12
- 238000001514 detection method Methods 0.000 claims description 11
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 10
- 239000007787 solid Substances 0.000 claims description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 9
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 8
- 238000004440 column chromatography Methods 0.000 claims description 7
- HZNVUJQVZSTENZ-UHFFFAOYSA-N 2,3-dichloro-5,6-dicyano-1,4-benzoquinone Chemical compound ClC1=C(Cl)C(=O)C(C#N)=C(C#N)C1=O HZNVUJQVZSTENZ-UHFFFAOYSA-N 0.000 claims description 6
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 6
- 239000003446 ligand Substances 0.000 claims description 5
- 229910052757 nitrogen Inorganic materials 0.000 claims description 5
- 238000011895 specific detection Methods 0.000 claims description 5
- 239000011259 mixed solution Substances 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 4
- 238000000967 suction filtration Methods 0.000 claims description 4
- 239000000839 emulsion Substances 0.000 claims description 3
- 239000012299 nitrogen atmosphere Substances 0.000 claims description 3
- 150000003904 phospholipids Chemical class 0.000 claims description 3
- 238000000108 ultra-filtration Methods 0.000 claims description 3
- 206010061218 Inflammation Diseases 0.000 claims description 2
- 238000001727 in vivo Methods 0.000 claims description 2
- 238000011503 in vivo imaging Methods 0.000 claims description 2
- 230000004054 inflammatory process Effects 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- 150000002503 iridium Chemical class 0.000 abstract 1
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 27
- WQYVRQLZKVEZGA-UHFFFAOYSA-N hypochlorite Chemical compound Cl[O-] WQYVRQLZKVEZGA-UHFFFAOYSA-N 0.000 description 26
- 238000012360 testing method Methods 0.000 description 17
- 238000005481 NMR spectroscopy Methods 0.000 description 8
- 239000000523 sample Substances 0.000 description 8
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 8
- 238000000862 absorption spectrum Methods 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 7
- 239000012327 Ruthenium complex Substances 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 5
- 238000003384 imaging method Methods 0.000 description 5
- 239000002904 solvent Substances 0.000 description 4
- 238000004448 titration Methods 0.000 description 4
- 102000003896 Myeloperoxidases Human genes 0.000 description 3
- 108090000235 Myeloperoxidases Proteins 0.000 description 3
- 239000005708 Sodium hypochlorite Substances 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000010226 confocal imaging Methods 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
- 230000003013 cytotoxicity Effects 0.000 description 3
- 239000007850 fluorescent dye Substances 0.000 description 3
- 230000003834 intracellular effect Effects 0.000 description 3
- 238000004020 luminiscence type Methods 0.000 description 3
- 230000001590 oxidative effect Effects 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 230000003595 spectral effect Effects 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000004847 absorption spectroscopy Methods 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000010949 copper Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000000295 emission spectrum Methods 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 230000036542 oxidative stress Effects 0.000 description 2
- 230000001766 physiological effect Effects 0.000 description 2
- 239000003642 reactive oxygen metabolite Substances 0.000 description 2
- 238000011897 real-time detection Methods 0.000 description 2
- 150000003303 ruthenium Chemical class 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010047115 Vasculitis Diseases 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 229910002091 carbon monoxide Inorganic materials 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 238000002784 cytotoxicity assay Methods 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000002296 dynamic light scattering Methods 0.000 description 1
- 238000004993 emission spectroscopy Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- CUILPNURFADTPE-UHFFFAOYSA-N hypobromous acid Chemical class BrO CUILPNURFADTPE-UHFFFAOYSA-N 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000011246 intracellular protein detection Methods 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000008204 material by function Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 201000008383 nephritis Diseases 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012113 quantitative test Methods 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000011896 sensitive detection Methods 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000000451 tissue damage Effects 0.000 description 1
- 231100000827 tissue damage Toxicity 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229910052723 transition metal Inorganic materials 0.000 description 1
- 150000003624 transition metals Chemical class 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 238000001392 ultraviolet--visible--near infrared spectroscopy Methods 0.000 description 1
- 229920003169 water-soluble polymer Polymers 0.000 description 1
Images
Classifications
-
- 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
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F15/00—Compounds containing elements of Groups 8, 9, 10 or 18 of the Periodic Table
- C07F15/0006—Compounds containing elements of Groups 8, 9, 10 or 18 of the Periodic Table compounds of the platinum group
- C07F15/0033—Iridium compounds
-
- 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
-
- 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
- C09K2211/18—Metal complexes
- C09K2211/185—Metal complexes of the platinum group, i.e. Os, Ir, Pt, Ru, Rh or Pd
Definitions
- the invention belongs to the technical field of organic photoelectric functional materials, and particularly relates to a water-soluble phosphorescent nano particle which can be used for detecting hypochlorous acid by a ratio method, a preparation method thereof and an application of the nano particle to detect hypochlorous acid in the living body.
- Active oxygen is a general term for a series of chemically active and oxidizing oxygen-containing substances produced by organisms.
- Reactive oxygen contains both free radicals and some non-free radicals such as hydrogen peroxide (H 2 O 2 ), hypochlorous acid (HClO), hydroxyl radicals (HO ⁇ ) and singlet oxygen ( 1 O 2 ).
- Reactive oxygen species play an important role in biological systems.
- HClO is a relatively common reactive oxygen species.
- the endogenous HClO is mediated by myeloperoxidase (MPO).
- MPO myeloperoxidase
- ClO - has high reactivity, short life span and more physiological activity. The process is an important and powerful oxidant that exerts an anti-microbial effect under physiological conditions and protects the body. Studies have shown that HClO is also a natural adaptive immune adjuvant. However, under certain conditions, if the excess of HClO produced by the MPO catalytic reaction exceeds the defense reaction of the local antioxidant, it will cause oxidative stress and oxidative tissue damage.
- Oxidative stress caused by excessive HClO has been shown to be associated with various diseases such as leukemia, nephritis, small vasculitis, tumors and atherosclerosis. Therefore, rapid, sensitive and real-time detection of hypochlorous acid has important physiological and pathological effects, which can provide reliable information for the pathogenesis, diagnosis and intervention of diseases.
- Phosphorescent transition metal complexes such as Pt(II)-, Ir(III)-, Ru(II)-, Cu(I)-, Au(I)-, etc., have been used in the field of living cell imaging in recent years. To gradually attract people's attention.
- the ruthenium complex exhibits special photoelectric properties in charge transfer and energy transfer between the metal center and the ligand, and has the advantages of high-efficiency triplet phosphorescence emission, long life, and large Stokes shift. It has not been found to be significantly toxic to cells and has great potential for application in cell biology imaging.
- the present invention aims to provide a water-soluble phosphorescent nanoparticle capable of detecting hypochlorous acid by a ratio method and to disclose a preparation method thereof and related applications, and the ratio method can reduce interference factors and accurately and specifically detect hypochlorous acid.
- the nanoparticle has excellent water solubility and biocompatibility, and has good application prospects in the fields of intracellular detection and living body detection.
- a water-soluble phosphorescent nanoparticle for detecting hypochlorous acid by a ratio method characterized in that the structure is as follows:
- C ⁇ N ligand in Ir(1-9) is any one of the following:
- the N ⁇ N ligand in Ir(1-6) * is any one of the following:
- water-soluble phosphorescent nanoparticles can be used for specific detection of hypochlorous acid by a ratio method.
- water-soluble phosphorescent nanoparticles can be used in the field of cell sensing and in vivo imaging sensing.
- water soluble phosphorescent nanoparticles can be used to establish a model of in vivo inflammation.
- the invention has the beneficial effects that the present invention prepares a water-soluble phosphorescent nanoparticle by solving the problems of poor biocompatibility, poor water solubility, short life span and high toxicity of the small molecule hypochlorous acid probe in the prior art.
- the nanoparticle binding ratio method realizes self-calibration, and can specifically detect the change of hypochlorous acid content; has a long emission life, combines time-resolved technology to eliminate interference of background fluorescent signal, and improves detection signal-to-noise ratio; meanwhile, the nanoparticle has Good water solubility and biocompatibility, can detect intracellular hypochlorous acid; low toxicity, less damage to biological samples, can achieve detection of hypochlorous acid in the living field.
- Example 1 is a responsive ultraviolet absorption spectrum of ruthenium complex Ir1 to hypochlorite in Example 4 of the present invention
- Figure 2 is a responsive ultraviolet absorption spectrum of ruthenium complex Ir1* to hypochlorite in Example 4 of the present invention
- Figure 3 is the embodiment Ir1-ClO 5 * Ir1 embodiment of the present invention, hypochlorous acid in response - emission spectrum;
- Example 6 is a statistical diagram showing ion exchange experiment results of Ir1 and Ir1* in Example 6 of the present invention
- Example 7 is a TEM test chart of phosphorescent water-soluble nanoparticle Ir NPs in Example 7 of the present invention.
- Example 6 is a DLS test chart of phosphorescent water-soluble nanoparticle Ir NPs in Example 8 of the present invention.
- Figure 7 is an absorption spectrum diagram of the complexes Ir1, Ir1* and the nanoparticles Ir NPs in Example 9 of the present invention.
- Figure 8 is a graph showing the titration spectrum of phosphorescent water-soluble nanoparticle Ir NPs in Example 10 of the present invention.
- Figure 9 is a test chart showing the relationship between the ratio of two emission peaks (I 600 /I 680 ) in the titration emission spectrum of the phosphorescent water-soluble nanoparticle Ir NPs according to the concentration of NaClO in the tenth embodiment of the present invention
- Figure 10 is a graph showing the experimental results of MTT cytotoxicity of phosphorescent water-soluble nanoparticle Ir NPs in Example 11 of the present invention.
- Figure 11 is a cell confocal imaging map of phosphorescent water-soluble nanoparticle Ir NPs in Example 12 of the present invention.
- the chemical reagents used in the present invention are all commercially available.
- the instruments used include:
- UV spectrometer UV-3600 UV-VIS-NIR, Shimadzu
- the probe complex Ir1 (1.0 mg) and the reference complex Ir1* (0.9 mg) were dissolved in a certain amount of tetrahydrofuran (2.0 mL), and a solution of 10.0 mg of phospholipid polyethylene glycol in PBS (10.0 mL) was added. Mix quickly and sonicate for 2.0 min. Then, the mixture was purged with a nitrogen balloon to tetrahydrofuran, and finally centrifuged in an ultrafiltration centrifuge tube to obtain an orange-red emulsion product, which was lyophilized to obtain an orange-red solid, that is, a nanoparticle Ir NPs.
- the ruthenium complex Ir1, Ir1* used in the present invention has a spectral test concentration of 10 ⁇ M, and the test solvent is a PBS solution mixed with 1% DMSO.
- Figure 1 is a UV absorption spectrum of the probe complex Ir1 after adding different concentrations of hypochlorite.
- Example 5 Responsive emission spectroscopy of hypobromite complexes Ir1 and Ir1*
- the ruthenium complex Ir1, Ir1* used in the present invention has a spectral test concentration of 10 ⁇ M, and the test solvent is a PBS solution mixed with 1% DMSO.
- the hypochlorite was 5 times the equivalent concentration, the response time was 1 minute, the remaining ions were 20 times the equivalent concentration, and the response time was 5 minutes.
- the results are shown in Fig. 4.
- the intensity of the emission peak of the reference complex Ir1* at 680 nm is almost constant, and the emission of the probe complex Ir1 at 680 nm is obtained.
- the peak is specifically illuminated by hypochlorite. Therefore, Ir1* can be used as a reference and imaged with the Ir1 construction ratio method for specific detection of changes in hypochlorite.
- Example 7 TEM test of phosphorescent water-soluble nanoparticle Ir NPs
- the phosphorescent water-soluble nanoparticle Ir NPs was dissolved in ethanol, dropped on a copper mesh, and subjected to TEM test after being naturally volatilized. As a result, as shown in Fig. 5, the nanoparticles were regular in shape, uniform in distribution, and all of them were circular, and the particle radius was about 105 nm.
- Example 8 DLS test of phosphorescent water-soluble nanoparticle Ir NPs
- the phosphorescent water-soluble nanoparticle Ir NPs was dissolved in ultrapure water, and the bubbles were removed by ultrasonication, and the DLS test was performed. As a result, as shown in Fig. 6, the nanoparticles were concentrated and had a hydration kinetic radius of about 125 nm.
- Example 9 Absorption spectroscopy test of complex Ir1, Ir1* and nanoparticle Ir NPs
- the ruthenium complexes Ir1 and Ir1* used in the present invention have a spectral concentration of 10 ⁇ M
- the test solvent is a PBS solution mixed with 1% DMSO
- the nanoparticles Ir NPs is 1 mg/mL
- the test solvent is a PBS solution.
- the ultraviolet absorption spectrum of the nanoparticle Ir NPs contains characteristic absorption peaks of the ruthenium complexes Ir1 and Ir1*.
- Example 11 MTT cytotoxicity assay of phosphorescent water-soluble nanoparticle Ir NPs
- the digested cells were seeded in a 96-well plate at a seeding density of 10 4 cells/well per well, and culture was continued for 24 hours at 37 ° C under 5% CO 2 . After the old culture solution was aspirated, the cells were further cultured for 24 hours with cell culture solutions of different concentrations of Ir NPs (10, 50, 100, 200, 300 ⁇ g/mL). The culture was terminated by adding 10 ⁇ L of MTT (5 mg/mL) to each well and continuing the culture for 4 hours. The culture solution was aspirated, 150 ⁇ L of DMSO was added to each well, and the shaker was shaken for 10 minutes, and then the OD570 was tested using a microplate reader.
- Fig. 10 The results of the MTT cytotoxicity experiment are shown in Fig. 10. It can be seen from the figure that when the concentration of the complex is 10 to 300 ⁇ g/mL, the cell survival rate after 24 hours of culture is greater than 90%, which proves that the nanoparticles have a lower concentration. Cytotoxicity can be used for cell imaging.
- Example 12 Confocal imaging experiment of phosphorescent water-soluble nanoparticle Ir NPs
- the cell confocal imaging experiment of the nanoparticles Ir NPs the experimental results are shown in Figure 11, using a concentration of 10 ⁇ g / mL.
- the specific procedure was to incubate HeLa cells in a 37 ° C incubator for 24 hours, then incubate HeLa cells with nanoparticle Ir NPs for 1 hour at 37 ° C, and then incubate with different concentrations of hypochlorous acid culture. Confocal testing was performed at the end of the incubation. The test results are shown in Fig. 11. The luminescence of the green channel increases with the increase of sodium hypochlorite concentration, while the luminescence of the red channel does not change significantly.
- the ratio of I 600nm /I 680nm increases with the increase of sodium hypochlorite. It is therefore possible to monitor changes in intracellular sodium hypochlorite by monitoring the luminescence and the ratio of the two. It is indicated that the phosphorescent water-soluble nanoparticle Ir NPs can specifically detect intracellular hypochlorous acid by ratiometric method combined with cell life imaging.
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Optics & Photonics (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
Provided is a water-soluble phosphorescent nanoparticle for detecting hypochlorous acid using a ratio method, consisting of iridium complexes Ir(1‑9) and Ir(1‑6)* and phospholipid-polyethylene glycol. The Ir(1‑9) can specifically respond to hypochlorous acid, and the Ir(1‑6)* is used as a reference complex and does not respond to hypochlorous acid. Also provided is a method for preparing a water-soluble phosphorescent nanoparticle.
Description
本发明属于有机光电功能材料技术领域，具体涉及一种可用于比率法检测次氯酸的水溶性磷光纳米粒子和它的制备方法以及该种纳米粒子在活体领域检测次氯酸上的应用。The invention belongs to the technical field of organic photoelectric functional materials, and particularly relates to a water-soluble phosphorescent nano particle which can be used for detecting hypochlorous acid by a ratio method, a preparation method thereof and an application of the nano particle to detect hypochlorous acid in the living body.
活性氧是生物体产生的一系列化学性质活泼、氧化能力强的含氧物质的总称。活性氧既包含自由基也包含一些非自由基，如过氧化氢(H
2O
2)，次氯酸(HClO)，羟自由基(HO·)和单线态氧(
1O
2)等，这些活性氧物质在生物***中扮演重要的角色。
Active oxygen is a general term for a series of chemically active and oxidizing oxygen-containing substances produced by organisms. Reactive oxygen contains both free radicals and some non-free radicals such as hydrogen peroxide (H 2 O 2 ), hypochlorous acid (HClO), hydroxyl radicals (HO·) and singlet oxygen ( 1 O 2 ). Reactive oxygen species play an important role in biological systems.
其中，HClO是一种较为常见的活性氧物种，生物体内源性HClO由髓过氧化物酶(MPO)介导产生，ClO
-具有较高的反应性，较短的寿命，参与较多的生理过程，是一种重要的功能强大的氧化剂，在生理状态下发挥抗微生物效应，起到保护机体的作用。研究表明，HClO还是天然的适应性免疫佐剂。然而，在特定条件下，假如MPO催化反应所产生的HClO过量，超过局部抗氧化剂的防御反应时，将导致氧化应激和氧化性组织损伤。已证实，过量HClO引起的氧化应激与白血病、肾炎、小血管炎、肿瘤及动脉粥样硬化等多种疾病相关。因此，快速、灵敏、实时检测次氯酸具有重要的生理、病理作用，可为疾病的发病机制、诊断及干预的研究提供可靠的信息。
Among them, HClO is a relatively common reactive oxygen species. The endogenous HClO is mediated by myeloperoxidase (MPO). ClO - has high reactivity, short life span and more physiological activity. The process is an important and powerful oxidant that exerts an anti-microbial effect under physiological conditions and protects the body. Studies have shown that HClO is also a natural adaptive immune adjuvant. However, under certain conditions, if the excess of HClO produced by the MPO catalytic reaction exceeds the defense reaction of the local antioxidant, it will cause oxidative stress and oxidative tissue damage. Oxidative stress caused by excessive HClO has been shown to be associated with various diseases such as leukemia, nephritis, small vasculitis, tumors and atherosclerosis. Therefore, rapid, sensitive and real-time detection of hypochlorous acid has important physiological and pathological effects, which can provide reliable information for the pathogenesis, diagnosis and intervention of diseases.
目前所报导的检测次氯酸的方法大多是利用小分子荧光探针，荧光探针在应用中大都基于单波长发射的荧光信号变化(通常是增强)，虽然探针能够实现对次氯酸的检测，但是大多数荧光探针存在各种各样的问题：量子效率低、水溶性差、易受背景荧光的干扰、准确性差等，无法实现对次氯酸的特异性、实时检测。而磷光过渡金属配合物，如Pt(II)-、Ir(III)-、Ru(II)-、Cu(I)-、Au(I)-等配合物，在活细胞成像领域的应用在近年来逐渐引起人们的关注。其中的铱配合物，在金属中心和配体之间的电荷转移和能量迁移展现出特殊的光电性能，具有高效的三线态磷光发射，较长的寿命，大的斯托克斯位移等优点，未发现对细胞有明显毒性，在细胞生物学成像方面有较大的应用潜力。Most of the methods reported for the detection of hypochlorous acid currently use small-molecule fluorescent probes, which are mostly based on single-wavelength emission of fluorescent signal changes (usually enhanced), although the probe can achieve hypochlorous acid. Detection, but most fluorescent probes have various problems: low quantum efficiency, poor water solubility, interference with background fluorescence, poor accuracy, etc., and the specific and real-time detection of hypochlorous acid cannot be achieved. Phosphorescent transition metal complexes, such as Pt(II)-, Ir(III)-, Ru(II)-, Cu(I)-, Au(I)-, etc., have been used in the field of living cell imaging in recent years. To gradually attract people's attention. Among them, the ruthenium complex exhibits special photoelectric properties in charge transfer and energy transfer between the metal center and the ligand, and has the advantages of high-efficiency triplet phosphorescence emission, long life, and large Stokes shift. It has not been found to be significantly toxic to cells and has great potential for application in cell biology imaging.
目前，关于以铱配合物为检测位点，水溶性聚合物为包覆材料的用于检测次 氯酸的水溶性磷光纳米粒子的报道还较为少见；且常见的次氯酸探针多为小分子荧光探针，生物相容性差，寿命短，量子效率低，而大多数生物成像重金属配合物的水溶性差，不利于生物体内次氯酸的特异性检测。同时，在现有技术中，通常是利用一个信息通道内的发射强度的变化来指示次氯酸的含量，这种方法很难在微环境中实现准确的定量测量。因此，如果我们要得到细胞微环境中被分析物的定量信息，就需要建立一套比率法，也就是让每个磷光纳米粒子在分子水平上建立内部的标尺，使之都具有自动校准功能。这意味着需要在原有的发射波长信号通道以外，引入第二个发射波长信号通道,通过双波长信号的比值的测量，减少或消除若干因素，如背景荧光和探针浓度等的变化对测量的影响，实现自校准，从而获得准确定量的信息。At present, reports on the use of water-soluble polymers as coating materials for the detection of water-soluble phosphorescent nanoparticles of hypochlorous acid with ruthenium complex as detection site are rare; and common hypochlorous acid probes are mostly small. Molecular fluorescent probes have poor biocompatibility, short lifetime and low quantum efficiency, while most bioimaging heavy metal complexes have poor water solubility, which is not conducive to the specific detection of hypochlorous acid in organisms. At the same time, in the prior art, it is common to utilize a change in emission intensity within an information channel to indicate the level of hypochlorous acid, which is difficult to achieve accurate quantitative measurements in a microenvironment. Therefore, if we want to obtain quantitative information about the analyte in the cell microenvironment, we need to establish a ratio method, that is, let each phosphorescent nanoparticle establish an internal scale at the molecular level, so that it has an automatic calibration function. This means that a second transmit wavelength signal path needs to be introduced outside the original transmit wavelength signal path, and the ratio of the dual-wavelength signal is measured to reduce or eliminate several factors such as background fluorescence and probe concentration. Affect, self-calibration to obtain accurate quantitative information.
因此，设计合成一种用于比率法检测次氯酸的水溶性磷光纳米粒子是非常有必要的。Therefore, it is very necessary to design and synthesize a water-soluble phosphorescent nanoparticle for the ratiometric detection of hypochlorous acid.
发明内容Summary of the invention
针对上述存在的问题，本发明旨在提供一种可用比率法检测次氯酸的水溶性磷光纳米粒子并公开其制备方法和相关应用，比率法可减少干扰因素进而精确、特异性检测次氯酸，且该纳米粒子具有优良的水溶性和生物相容性，在胞内检测和活体检测等领域具有良好的应用前景。In view of the above problems, the present invention aims to provide a water-soluble phosphorescent nanoparticle capable of detecting hypochlorous acid by a ratio method and to disclose a preparation method thereof and related applications, and the ratio method can reduce interference factors and accurately and specifically detect hypochlorous acid. The nanoparticle has excellent water solubility and biocompatibility, and has good application prospects in the fields of intracellular detection and living body detection.
为了实现上述目的，本发明所采用的技术方案如下：一种用于比率法检测次氯酸的水溶性磷光纳米粒子，其特征在于，其结构如下：In order to achieve the above object, the technical solution adopted by the present invention is as follows: a water-soluble phosphorescent nanoparticle for detecting hypochlorous acid by a ratio method, characterized in that the structure is as follows:
该水溶性磷光纳米粒子的具体合成路线为：The specific synthetic route of the water-soluble phosphorescent nanoparticle is:
其中，Ir(1-9)中的C^N配体为下列中的任一个：Wherein the C^N ligand in Ir(1-9) is any one of the following:
Ir(1-6)
*中的N^N配体为下列中的任一个：
The N^N ligand in Ir(1-6) * is any one of the following:
所述水溶性磷光纳米粒子的制备方法的具体操作步骤如下：The specific operation steps of the preparation method of the water-soluble phosphorescent nano particles are as follows:
1)将化合物a、化合物b和碳酸钾，在氮气保护下在乙腈溶液中反应3-5h，过滤，柱层析分离得到橙黄色固体化合物c；1) The compound a, the compound b and the potassium carbonate are reacted in an acetonitrile solution under a nitrogen atmosphere for 3-5 hours, filtered, and separated by column chromatography to obtain an orange-yellow solid compound c;
2)将所述化合物c、铱二氯桥和六氟磷酸钾，在氮气保护下溶解于二氯甲烷和甲醇的混合溶液中，于45℃条件下回流反应6h，抽滤，柱层析分离得到橙红色固体，即配合物Ir(1-9)；2) The compound c, bismuth dichloride bridge and potassium hexafluorophosphate are dissolved in a mixed solution of dichloromethane and methanol under nitrogen protection, refluxed at 45 ° C for 6 h, suction filtration, column chromatography separation Obtaining an orange-red solid, ie complex Ir (1-9);
3)将所述配合物Ir(1-9)和配合物Ir(1-6)
*溶解于四氢呋喃中，加入磷脂聚乙二醇的PBS溶液，迅速混合并超声2.0min，氮气球鼓气至四氢呋喃挥发，用超滤离心管离心得到橙红色乳液产物，经冻干得到橙红色固体，即纳米粒子Ir NPs。
3) The complex Ir(1-9) and the complex Ir(1-6) * were dissolved in tetrahydrofuran, and a solution of phospholipid polyethylene glycol in PBS was added, and the mixture was rapidly mixed and ultrasonicated for 2.0 min. The tetrahydrofuran was volatilized and centrifuged in an ultrafiltration centrifuge tube to obtain an orange-red emulsion product, which was lyophilized to obtain an orange-red solid, i.e., a nanoparticle Ir NPs.
进一步地，所述水溶性磷光纳米粒子可用于比率法特异性检测次氯酸。Further, the water-soluble phosphorescent nanoparticles can be used for specific detection of hypochlorous acid by a ratio method.
进一步地，所述水溶性磷光纳米粒子可用于细胞传感和活体成像传感领域。Further, the water-soluble phosphorescent nanoparticles can be used in the field of cell sensing and in vivo imaging sensing.
进一步地，所述水溶性磷光纳米粒子可用于建立活体炎症模型。Further, the water soluble phosphorescent nanoparticles can be used to establish a model of in vivo inflammation.
本发明的有益效果是：本发明为了解决现有技术中的小分子次氯酸探针生物相容性差、水溶性差、寿命短、毒性大等多种问题制备了一种水溶性磷光纳米粒 子，该纳米粒子结合比率法实现自校准，能够特异性检测次氯酸含量的变化；具有长的发射寿命，结合时间分辨技术排除背景荧光信号的干扰，提高检测信噪比；同时，该纳米粒子具有良好的水溶性和生物相容性，可实现对细胞内次氯酸的检测；毒性小，对生物样品的损伤小，可以实现对活体领域次氯酸的检测。The invention has the beneficial effects that the present invention prepares a water-soluble phosphorescent nanoparticle by solving the problems of poor biocompatibility, poor water solubility, short life span and high toxicity of the small molecule hypochlorous acid probe in the prior art. The nanoparticle binding ratio method realizes self-calibration, and can specifically detect the change of hypochlorous acid content; has a long emission life, combines time-resolved technology to eliminate interference of background fluorescent signal, and improves detection signal-to-noise ratio; meanwhile, the nanoparticle has Good water solubility and biocompatibility, can detect intracellular hypochlorous acid; low toxicity, less damage to biological samples, can achieve detection of hypochlorous acid in the living field.
图1是本发明实施例4中铱配合物Ir1对次氯酸根的响应性紫外吸收光谱图；1 is a responsive ultraviolet absorption spectrum of ruthenium complex Ir1 to hypochlorite in Example 4 of the present invention;
图2是本发明实施例4中铱配合物Ir1*对次氯酸根的响应性紫外吸收光谱图；Figure 2 is a responsive ultraviolet absorption spectrum of ruthenium complex Ir1* to hypochlorite in Example 4 of the present invention;
图3是本发明实施例5中Ir1*、次氯酸响应后的Ir1-ClO
-的发射光谱图；
Figure 3 is the embodiment Ir1-ClO 5 * Ir1 embodiment of the present invention, hypochlorous acid in response - emission spectrum;
图4是本发明实施例6中Ir1、Ir1*的离子选择性实验结果统计图；4 is a statistical diagram showing ion exchange experiment results of Ir1 and Ir1* in Example 6 of the present invention;
图5是本发明实施例7中磷光水溶性纳米粒子Ir NPs的TEM测试图；5 is a TEM test chart of phosphorescent water-soluble nanoparticle Ir NPs in Example 7 of the present invention;
图6是本发明实施例8中磷光水溶性纳米粒子Ir NPs的DLS测试图；6 is a DLS test chart of phosphorescent water-soluble nanoparticle Ir NPs in Example 8 of the present invention;
图7是本发明实施例9中配合物Ir1、Ir1*和纳米粒子Ir NPs的吸收光谱图；Figure 7 is an absorption spectrum diagram of the complexes Ir1, Ir1* and the nanoparticles Ir NPs in Example 9 of the present invention;
图8是本发明实施例10中磷光水溶性纳米粒子Ir NPs的滴定光谱测试图；Figure 8 is a graph showing the titration spectrum of phosphorescent water-soluble nanoparticle Ir NPs in Example 10 of the present invention;
图9是本发明实施例10中磷光水溶性纳米粒子Ir NPs的滴定发射光谱中两个发射峰比值(I
600/I
680)的大小随着NaClO浓度变化的测试图；
Figure 9 is a test chart showing the relationship between the ratio of two emission peaks (I 600 /I 680 ) in the titration emission spectrum of the phosphorescent water-soluble nanoparticle Ir NPs according to the concentration of NaClO in the tenth embodiment of the present invention;
图10是本发明实施例11中磷光水溶性纳米粒子Ir NPs的MTT细胞毒性实验统计图；Figure 10 is a graph showing the experimental results of MTT cytotoxicity of phosphorescent water-soluble nanoparticle Ir NPs in Example 11 of the present invention;
图11是本发明实施例12中磷光水溶性纳米粒子Ir NPs的细胞共聚焦成像图谱。Figure 11 is a cell confocal imaging map of phosphorescent water-soluble nanoparticle Ir NPs in Example 12 of the present invention.
为了使本领域的普通技术人员能更好的理解本发明的技术方案，下面结合附图和实施例对本发明的技术方案做进一步的描述。In order to enable a person skilled in the art to better understand the technical solutions of the present invention, the technical solutions of the present invention will be further described below in conjunction with the accompanying drawings and embodiments.
本发明中使用的化学试剂皆为市购。The chemical reagents used in the present invention are all commercially available.
使用的仪器包括：The instruments used include:
发射光谱仪：Edinburgh FL 920，EdinburghEmission spectrometer: Edinburgh FL 920, Edinburgh
紫外光谱仪：UV-3600 UV-VIS-NIR，ShimadzuUV spectrometer: UV-3600 UV-VIS-NIR, Shimadzu
核磁共振：Ultra Shield Plus 400MHz NMR，BrukerNMR: Ultra Shield Plus 400MHz NMR, Bruker
透射电镜：JEOL JEM-2100，JEOLTransmission electron microscopy: JEOL JEM-2100, JEOL
动态光散射仪：，ZetasizerNanoseries，MalvernDynamic Light Scattering Apparatus:, ZetasizerNanoseries, Malvern
共聚焦扫描仪：Becker &Hickl GmbH DCS-120，Becker &Hickl GmbHConfocal Scanner: Becker &Hickl GmbH DCS-120, Becker &Hickl GmbH
实施例1：对次氯酸敏感的配合物Ir1的制备：Example 1: Preparation of hypochlorous acid-sensitive complex Ir1:
(1)化合物c的制备：将化合物a(1.5mmol)、化合物b(1.0mmol)和碳酸钾1(2.0mmol)在乙腈溶液(12mL)中于氮气保护下反应4.5h。反应结束后，过滤除去碳酸钾并通过柱层析分离得到橙黄色固体化合物c，产率70％；(1) Preparation of Compound c: Compound a (1.5 mmol), Compound b (1.0 mmol), and potassium carbonate 1 (2.0 mmol) were reacted in an acetonitrile solution (12 mL) under nitrogen for 4.5 h. After the reaction is completed, potassium carbonate is removed by filtration and separated by column chromatography to obtain an orange-yellow solid compound c in a yield of 70%;
1H NMR(400MHz,DMSO)δ(ppm):8.68(dd,J＝5.2Hz,9.2Hz,2H),8.45(d,J＝8.0Hz,2H),7.51(s,2H),7.48(d,J＝5.2Hz,2H),7.31(dd,J＝4.4Hz,8.0Hz,3H),7.01(d,J＝9.2Hz,1H),5.24(s,1H),4.78(s,1H),
13C NMR(100MHz,DMSO)δ(ppm):155.9,155.4,150.3,149.9,148.4,148.1,147.9,142.7,129.5,128.0,124.7,122.8,121.4,121.2,118.8,107.3,68.7,32.3.
1 H NMR (400MHz, DMSO) δ (ppm): 8.68 (dd, J = 5.2Hz, 9.2Hz, 2H), 8.45 (d, J = 8.0Hz, 2H), 7.51 (s, 2H), 7.48 (d , J = 5.2 Hz, 2H), 7.31 (dd, J = 4.4 Hz, 8.0 Hz, 3H), 7.01 (d, J = 9.2 Hz, 1H), 5.24 (s, 1H), 4.78 (s, 1H), 13 C NMR (100 MHz, DMSO) δ (ppm): 155.9, 155.4, 150.3, 149.9, 148.4, 148.1, 147.9, 142.7, 129.5, 128.0, 124.7, 122.8, 121.4, 121.2, 118.8, 107.3, 68.7, 32.3.
(2)配合物Ir1的制备：将所述化合物c(0.5mmol)、铱二氯桥(0.25mmol)和足量六氟磷酸钾，在氮气保护下于二氯甲烷和甲醇的混合溶液中于45℃条件下回流反应6h，反应结束后，抽滤除去六氟磷酸钾，通过柱层析分离得到橙红色固体，产率80％；(2) Preparation of complex Ir1: the compound c (0.5 mmol), ruthenium dichloride bridge (0.25 mmol) and sufficient potassium hexafluorophosphate were mixed under a nitrogen atmosphere in a mixed solution of dichloromethane and methanol. The reaction was refluxed at 45 ° C for 6 h. After the reaction was completed, potassium hexafluorophosphate was removed by suction filtration and separated by column chromatography to give an orange-red solid.
1H NMR(400MHz,DMSO)δ(ppm):8.72(d,J＝12.4Hz,1H),8.65(d,J＝8.8Hz,1H),8.43(t,J＝8.0Hz,2H),8.06-7.99(m,4H),7.90(dd,J＝8.0Hz,4.0Hz,2H),7.83(dd,J＝5.2Hz,1.2Hz,2H),7.69(t,J＝4.0Hz,2H),7.44-7.34(m,5H),7.26(dd,J＝8.0Hz,4.0Hz,1H),7.09(t,J＝8.0Hz,1H),7.03-6.92(m,4H),6.17-6.14(m,2H),5.31(s,2H),4.90-4.74(m,2H),
13C NMR(100MHz,DMSO)δ(ppm):166.1,166.0,156.1,156.0,155.5,155.4,155.3,155.2,151.6,151.3,151.0,149.1,149.0,148.7,148.0,147.9,147.5,142.9,141.4,140.6,133.7,133.0,131.8,131.7,129.4,128.0,126.5,126.4,126.2,124.3,122.7,121.3,118.2,107.8,68.1,43.6,31.4
1 H NMR (400MHz, DMSO) δ (ppm): 8.72 (d, J = 12.4Hz, 1H), 8.65 (d, J = 8.8Hz, 1H), 8.43 (t, J = 8.0Hz, 2H), 8.06 -7.99 (m, 4H), 7.90 (dd, J = 8.0 Hz, 4.0 Hz, 2H), 7.83 (dd, J = 5.2 Hz, 1.2 Hz, 2H), 7.69 (t, J = 4.0 Hz, 2H), 7.44-7.34 (m, 5H), 7.26 (dd, J = 8.0 Hz, 4.0 Hz, 1H), 7.09 (t, J = 8.0 Hz, 1H), 7.03-6.92 (m, 4H), 6.17-6.14 (m , 2H), 5.31 (s, 2H), 4.90-4.74 (m, 2H), 13 C NMR (100 MHz, DMSO) δ (ppm): 166.1, 166.0, 156.1, 156.0, 155.5, 155.4, 155.3, 155.2, 151.6 , 151.3, 151.0, 149.1, 149.0, 148.7, 148.0, 147.9, 147.5, 142.9, 141.4, 140.6, 133.7, 133.0, 131.8, 131.7, 129.4, 128.0, 126.5, 126.4, 126.2, 124.3, 122.7, 121.3, 118.2, 107.8 ,68.1,43.6,31.4
实施例2：对次氯酸不敏感的参比配合物Ir1*的制备：Example 2: Preparation of a reference complex Ir1* which is insensitive to hypochlorous acid:
配合物Ir1*的制备：将1*(0.5mmol)、铱二氯桥(0.25mmol)和足量六氟磷酸钾，在氮气保护下，于二氯甲烷和甲醇的混合溶液中于45℃条件下回流反应6h，反应结束后，抽滤除去六氟磷酸钾，通过柱层析分离得到红色固体，产率85％；Preparation of complex Ir1*: 1*(0.5mmol), bismuth dichloride bridge (0.25mmol) and sufficient potassium hexafluorophosphate under nitrogen protection in a mixed solution of dichloromethane and methanol at 45 ° C The reaction was refluxed for 6 h, and after completion of the reaction, potassium hexafluorophosphate was removed by suction filtration, and a red solid was obtained by column chromatography to give a yield of 85%;
1H NMR(400MHz,CD
3OD)δ(ppm):9.00(d,J＝12.0Hz,2H),8.78(s,2H),8.15(d,J＝8.0Hz,2H),8.03-7.94(m,6H),7.66(d,J＝4.0Hz,2H),7.58(dd,J＝10.0Hz,7.2Hz,4H),7.48(d,J＝5.2Hz,2H),7.22(t,J＝7.6Hz,2H),6.74(t,J＝7.6Hz,2H),6.06(d,J＝8.0Hz,2H).
13C NMR(100MHz,DMSO)δ(ppm):164.9,156.5,155.3,152.9,148.9,144.5,143.5,142.2,137.1,135.6,133.4,130.5,130.2,128.5,127.3,125.0,124.6,123.3,120.7,21.3.
1 H NMR (400 MHz, CD 3 OD) δ (ppm): 9.00 (d, J = 12.0 Hz, 2H), 8.78 (s, 2H), 8.15 (d, J = 8.0 Hz, 2H), 8.03 - 7.94 ( m,6H), 7.66 (d, J=4.0 Hz, 2H), 7.58 (dd, J = 10.0 Hz, 7.2 Hz, 4H), 7.48 (d, J = 5.2 Hz, 2H), 7.22 (t, J = 7.6 Hz, 2H), 6.74 (t, J = 7.6 Hz, 2H), 6.06 (d, J = 8.0 Hz, 2H). 13 C NMR (100 MHz, DMSO) δ (ppm): 164.9, 156.5, 155.3, 152.9 , 148.9, 144.5, 143.5, 142.2, 137.1, 135.6, 133.4, 130.5, 130.2, 128.5, 127.3, 125.0, 124.6, 123.3, 120.7, 21.3.
实施例3：对次氯酸敏感的水溶性磷光纳米粒子的制备：Example 3: Preparation of water-soluble phosphorescent nanoparticles sensitive to hypochlorous acid:
将探针配合物Ir1(1.0mg)和参比配合物Ir1*(0.9mg)溶解于一定量的四氢呋喃(2.0mL)中，加入含有10.0mg磷脂聚乙二醇的PBS(10.0mL)溶液，迅速混合并超声2.0min。然后用氮气球鼓气至四氢呋喃挥发，最后用超滤离心管离心得到橙红色乳液产物，经冻干可得到橙红色固体，即纳米粒子Ir NPs。The probe complex Ir1 (1.0 mg) and the reference complex Ir1* (0.9 mg) were dissolved in a certain amount of tetrahydrofuran (2.0 mL), and a solution of 10.0 mg of phospholipid polyethylene glycol in PBS (10.0 mL) was added. Mix quickly and sonicate for 2.0 min. Then, the mixture was purged with a nitrogen balloon to tetrahydrofuran, and finally centrifuged in an ultrafiltration centrifuge tube to obtain an orange-red emulsion product, which was lyophilized to obtain an orange-red solid, that is, a nanoparticle Ir NPs.
实施例4：配合物Ir1、Ir1*对次氯酸根的响应性紫外吸收光谱测试Example 4: Responsive UV absorption spectroscopy of complexes Ir1, Ir1* for hypochlorite
本发明采用的铱配合物Ir1、Ir1*光谱测试浓度为10μM，测试溶剂为混有1％DMSO的PBS溶液。图1为探针配合物Ir1在加入不同浓度的次氯酸根后的紫外吸收光谱图，如图所示，随着次氯酸根浓度的增加，Ir1在300nm处的吸收峰略有下降；图2为参比配合物Ir1*在加入不同浓度的次氯酸根后的紫外吸收光谱，如图所示，随着次氯酸根浓度的增加，Ir1*的紫外吸收光谱几乎不变，该结果从一定程度上说明Ir1在与次氯酸根作用后结构发生了改变，而Ir1*几乎不与次氯酸发生反应。The ruthenium complex Ir1, Ir1* used in the present invention has a spectral test concentration of 10 μM, and the test solvent is a PBS solution mixed with 1% DMSO. Figure 1 is a UV absorption spectrum of the probe complex Ir1 after adding different concentrations of hypochlorite. As shown in the figure, with the increase of hypochlorite concentration, the absorption peak of Ir1 at 300 nm decreases slightly; The UV absorption spectrum of the reference complex Ir1* after adding different concentrations of hypochlorite, as shown in the figure, with the increase of hypochlorite concentration, the ultraviolet absorption spectrum of Ir1* is almost unchanged, the result is from a certain extent It is indicated that the structure of Ir1 changes after interaction with hypochlorite, and Ir1* hardly reacts with hypochlorous acid.
实施例5：配合物Ir1、Ir1*对次氯酸根的响应性发射光谱测试Example 5: Responsive emission spectroscopy of hypobromite complexes Ir1 and Ir1*
本发明采用的铱配合物Ir1、Ir1*光谱测试浓度为10μM，测试溶剂为混有 1％DMSO的PBS溶液。如图3所示，参比配合物Ir1*的最高发射峰为680nm，探针配合物Ir1与次氯酸反应后的Ir1-ClO
-的最高发射峰为600nm，两者相距较远，影响较小，可以用于构建比率法成像。
The ruthenium complex Ir1, Ir1* used in the present invention has a spectral test concentration of 10 μM, and the test solvent is a PBS solution mixed with 1% DMSO. As shown, the reference complex Ir1 3 * maximum emission peak of 680 nm, a probe complex after reaction with hypochlorite Ir1 Ir1-ClO - maximum emission peak of 600 nm, both the far distance, the more impact Small, can be used to construct ratiometric imaging.
实施例6：配合物Ir1、Ir1*的离子选择性实验Example 6: Ion Selectivity Experiment of Complex Ir1, Ir1*
次氯酸根为5倍当量浓度，响应时间为1分钟，其余离子为20倍当量浓度，响应时间5分钟。结果如图4所示，在ClO
-、K
+、H
2O
2等存在下，参比配合物Ir1*在680nm处的发射峰强度几乎不变，而探针配合物Ir1在680nm处的发射峰被次氯酸根特异性点亮。因此，Ir1*可以用作参比，与Ir1构建比率法成像，用于特异性检测次氯酸根的变化。
The hypochlorite was 5 times the equivalent concentration, the response time was 1 minute, the remaining ions were 20 times the equivalent concentration, and the response time was 5 minutes. The results are shown in Fig. 4. In the presence of ClO - , K + , H 2 O 2 , etc., the intensity of the emission peak of the reference complex Ir1* at 680 nm is almost constant, and the emission of the probe complex Ir1 at 680 nm is obtained. The peak is specifically illuminated by hypochlorite. Therefore, Ir1* can be used as a reference and imaged with the Ir1 construction ratio method for specific detection of changes in hypochlorite.
实施例7：磷光水溶性纳米粒子Ir NPs的TEM测试Example 7: TEM test of phosphorescent water-soluble nanoparticle Ir NPs
将磷光水溶性纳米粒子Ir NPs溶于乙醇中，滴在铜网上，待其自然挥发后进行TEM测试。结果如图5所示，纳米粒子形状规则，分布均匀，均为圆形，粒子半径约为105nm。The phosphorescent water-soluble nanoparticle Ir NPs was dissolved in ethanol, dropped on a copper mesh, and subjected to TEM test after being naturally volatilized. As a result, as shown in Fig. 5, the nanoparticles were regular in shape, uniform in distribution, and all of them were circular, and the particle radius was about 105 nm.
实施例8：磷光水溶性纳米粒子Ir NPs的DLS测试Example 8: DLS test of phosphorescent water-soluble nanoparticle Ir NPs
将磷光水溶性纳米粒子Ir NPs溶于超纯水中，超声除去气泡，进行DLS测试。结果如图6所示，纳米粒子集中分布，其水合动力学半径约为125nm。The phosphorescent water-soluble nanoparticle Ir NPs was dissolved in ultrapure water, and the bubbles were removed by ultrasonication, and the DLS test was performed. As a result, as shown in Fig. 6, the nanoparticles were concentrated and had a hydration kinetic radius of about 125 nm.
实施例9：配合物Ir1、Ir1*和纳米粒子Ir NPs的吸收光谱测试Example 9: Absorption spectroscopy test of complex Ir1, Ir1* and nanoparticle Ir NPs
本发明采用的铱配合物Ir1、Ir1*光谱测试浓度为10μM，测试溶剂为混有1％DMSO的PBS溶液；纳米粒子Ir NPs为1mg/mL，测试溶剂为PBS溶液。结果如图7所示，纳米粒子Ir NPs的紫外吸收光谱包含了铱配合物Ir1、Ir1*的特征吸收峰。The ruthenium complexes Ir1 and Ir1* used in the present invention have a spectral concentration of 10 μM, the test solvent is a PBS solution mixed with 1% DMSO, the nanoparticles Ir NPs is 1 mg/mL, and the test solvent is a PBS solution. As a result, as shown in FIG. 7, the ultraviolet absorption spectrum of the nanoparticle Ir NPs contains characteristic absorption peaks of the ruthenium complexes Ir1 and Ir1*.
实施例10：磷光水溶性纳米粒子Ir NPs的滴定光谱测试Example 10: Titration Spectrometric Test of Phosphorescent Water-Soluble Nanoparticle Ir NPs
纳米粒子Ir NPs与0-20μMNaClO在PBS溶液(pH＝7.4)中的滴定光谱测试结果如图8所示，随着NaClO的增加，600nm处的发射强度不断增加，而680nm处的发射强度变化很小。I
600nm/I
680nm与NaClO的关系如图9所示，从图中可以看出，随着NaClO的浓度不断增加，其比值不断增大，呈一定线性关系，可实现对次氯酸根的定量测试。
The results of titration spectroscopy of nanoparticle Ir NPs with 0-20 μM NaClO in PBS solution (pH=7.4) are shown in Fig. 8. With the increase of NaClO, the emission intensity at 600 nm increases continuously, and the emission intensity at 680 nm changes very much. small. The relationship between I 600nm /I 680nm and NaClO is shown in Fig. 9. It can be seen from the figure that as the concentration of NaClO increases, the ratio increases continuously and shows a linear relationship, which can realize the quantitative test of hypochlorite. .
实施例11：磷光水溶性纳米粒子Ir NPs的MTT细胞毒性实验Example 11: MTT cytotoxicity assay of phosphorescent water-soluble nanoparticle Ir NPs
将消化后的细胞接种在96孔板中，每孔的接种密度为10
4个/孔，在37℃，5％CO
2的条件下继续培养24小时。吸除旧的培养液后用不同浓度Ir NPs(10、 50、100、200、300μg/mL)的细胞培养液继续培养细胞24小时。每孔加入10μL MTT(5mg/mL)继续培养4小时后终止培养。吸除培养液，每孔加入150μL DMSO，摇床震荡10分钟后使用酶标仪测试OD570。
The digested cells were seeded in a 96-well plate at a seeding density of 10 4 cells/well per well, and culture was continued for 24 hours at 37 ° C under 5% CO 2 . After the old culture solution was aspirated, the cells were further cultured for 24 hours with cell culture solutions of different concentrations of Ir NPs (10, 50, 100, 200, 300 μg/mL). The culture was terminated by adding 10 μL of MTT (5 mg/mL) to each well and continuing the culture for 4 hours. The culture solution was aspirated, 150 μL of DMSO was added to each well, and the shaker was shaken for 10 minutes, and then the OD570 was tested using a microplate reader.
MTT细胞毒性实验结果如图10所示，从图中可以看出在配合物的浓度为10～300μg/mL时，培养24小时后的细胞存活率均大于90％，证明该纳米粒子具有较低的细胞毒性，可用于细胞成像。The results of the MTT cytotoxicity experiment are shown in Fig. 10. It can be seen from the figure that when the concentration of the complex is 10 to 300 μg/mL, the cell survival rate after 24 hours of culture is greater than 90%, which proves that the nanoparticles have a lower concentration. Cytotoxicity can be used for cell imaging.
实施例12：磷光水溶性纳米粒子Ir NPs的细胞共聚焦成像实验Example 12: Confocal imaging experiment of phosphorescent water-soluble nanoparticle Ir NPs
纳米粒子Ir NPs的细胞共聚焦成像实验，实验结果如图11所示，采用的浓度为10μg/mL。具体过程为，在37℃恒温箱中培养HeLa细胞24小时，然后将HeLa细胞与纳米粒子Ir NPs在37℃下共孵育1小时，然后采用不同浓度的次氯酸培养液孵育。孵育结束进行共聚焦测试。测试结果如图11所示，绿光通道的发光随着次氯酸钠浓度的升高而增强，而红光通道的发光无明显变化，其比值I
600nm/I
680nm随着次氯酸钠的升高而增加。因此可以通过监测发光和两者的比值来监测细胞内次氯酸钠的变化。说明磷光水溶性纳米粒子Ir NPs可以通过比率法结合细胞寿命成像来特异性检测细胞内的次氯酸。
The cell confocal imaging experiment of the nanoparticles Ir NPs, the experimental results are shown in Figure 11, using a concentration of 10 μg / mL. The specific procedure was to incubate HeLa cells in a 37 ° C incubator for 24 hours, then incubate HeLa cells with nanoparticle Ir NPs for 1 hour at 37 ° C, and then incubate with different concentrations of hypochlorous acid culture. Confocal testing was performed at the end of the incubation. The test results are shown in Fig. 11. The luminescence of the green channel increases with the increase of sodium hypochlorite concentration, while the luminescence of the red channel does not change significantly. The ratio of I 600nm /I 680nm increases with the increase of sodium hypochlorite. It is therefore possible to monitor changes in intracellular sodium hypochlorite by monitoring the luminescence and the ratio of the two. It is indicated that the phosphorescent water-soluble nanoparticle Ir NPs can specifically detect intracellular hypochlorous acid by ratiometric method combined with cell life imaging.
以上显示和描述了本发明的基本原理、主要特征及优点。但是以上所述仅为本发明的具体实施例，本发明的技术特征并不局限于此，任何本领域的技术人员在不脱离本发明的技术方案下得出的其他实施方式均应涵盖在本发明的专利范围之中。The basic principles, main features and advantages of the present invention have been shown and described above. However, the above description is only a specific embodiment of the present invention, and the technical features of the present invention are not limited thereto, and any other embodiments obtained by those skilled in the art without departing from the technical solution of the present invention should be covered in the present invention. Within the scope of the invention patent.
Claims (6)
- 一种用于比率法检测次氯酸的水溶性磷光纳米粒子，其特征在于，其结构如下：A water-soluble phosphorescent nanoparticle for detecting hypochlorous acid by a ratio method, characterized in that the structure is as follows:
- 如权利要求1所述的一种用于比率法检测次氯酸的水溶性磷光纳米粒子，其特征在于，其合成路线为：A water-soluble phosphorescent nanoparticle for detecting hypochlorous acid by a ratio method according to claim 1, wherein the synthetic route is:
其中，Ir(1-9)中的C^N配体为下列中的任一个：Wherein the C^N ligand in Ir(1-9) is any one of the following:
Ir(1-6) *中的N^N配体为下列中的任一个： The N^N ligand in Ir(1-6) * is any one of the following:
- 如权利要求1所述的一种用于比率法检测次氯酸的水溶性磷光纳米粒子的制备方法，其特征在于，具体操作步骤如下：The method for preparing a water-soluble phosphorescent nanoparticle for detecting hypochlorous acid by a ratio method according to claim 1, wherein the specific operation steps are as follows:1)将化合物a、化合物b和碳酸钾，在氮气保护下在乙腈溶液中混合反应3-5 h，过滤,柱层析分离得到橙黄色固体化合物c；1) Compound a, compound b and potassium carbonate are mixed and reacted in an acetonitrile solution under a nitrogen atmosphere for 3-5 h, filtered, and separated by column chromatography to obtain an orange-yellow solid compound c;2)将所述化合物c、铱二氯桥和六氟磷酸钾，在氮气保护下溶解于二氯甲烷和甲醇的混合溶液中，在45℃条件下回流反应6h，抽滤，柱层析分离得到橙红色固体，即配合物Ir(1-9)；2) The compound c, bismuth dichloride bridge and potassium hexafluorophosphate are dissolved in a mixed solution of dichloromethane and methanol under the protection of nitrogen, and refluxed at 45 ° C for 6 h, suction filtration, column chromatography separation Obtaining an orange-red solid, ie complex Ir (1-9);3)将所述配合物Ir(1-9)和配合物Ir(1-6) *溶解于四氢呋喃中，加入磷脂聚乙二醇的PBS溶液，迅速混合并超声2.0min，氮气球鼓气至四氢呋喃挥发，用超滤离心管离心得到橙红色乳液产物，经冻干得到橙红色固体，即纳米粒子Ir NPs。 3) The complex Ir(1-9) and the complex Ir(1-6) * were dissolved in tetrahydrofuran, and a solution of phospholipid polyethylene glycol in PBS was added, and the mixture was rapidly mixed and ultrasonicated for 2.0 min. The tetrahydrofuran was volatilized and centrifuged in an ultrafiltration centrifuge tube to obtain an orange-red emulsion product, which was lyophilized to obtain an orange-red solid, i.e., a nanoparticle Ir NPs.
- 如权利要求1-3中任一项所述的一种用于比率法检测次氯酸的水溶性磷光纳米粒子在比率法特异性检测次氯酸中的应用。Use of a water-soluble phosphorescent nanoparticle for ratiometric detection of hypochlorous acid according to any one of claims 1 to 3 for the specific detection of hypochlorous acid by a ratio method.
- 如权利要求1-3中任一项所述的一种用于比率法检测次氯酸的水溶性磷光纳米粒子在细胞传感领域和活体成像传感领域中的应用。Use of a water-soluble phosphorescent nanoparticle for ratiometric detection of hypochlorous acid according to any one of claims 1 to 3 in the field of cell sensing and in vivo imaging sensing.
- 如权利要求1-3中任一项所述的一种用于比率法检测次氯酸的水溶性磷光纳米粒子在活体炎症模型中的应用。Use of a water-soluble phosphorescent nanoparticle for ratiometric detection of hypochlorous acid according to any one of claims 1 to 3 in a model of in vivo inflammation.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810473705.3 | 2018-05-17 | ||
| CN201810473705.3A CN108535233B (en) | 2018-05-17 | 2018-05-17 | Water-soluble phosphorescent nanoparticles for detecting hypochlorous acid by ratio method and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2019218594A1 true WO2019218594A1 (en) | 2019-11-21 |
Family
ID=63472053
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/CN2018/111540 WO2019218594A1 (en) | 2018-05-17 | 2018-10-24 | Water-soluble phosphorescent nanoparticle for detecting hypochlorous acid using ratio method and preparation method and application thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN108535233B (en) |
| WO (1) | WO2019218594A1 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108535233B (en) * | 2018-05-17 | 2020-07-17 | 南京邮电大学 | Water-soluble phosphorescent nanoparticles for detecting hypochlorous acid by ratio method and preparation method and application thereof |
| CN111233754B (en) * | 2020-01-19 | 2023-06-02 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Phosphorescent probe based on platinum complex and application thereof in hypochlorous acid detection |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007100061A1 (en) * | 2006-03-03 | 2007-09-07 | The University Of Tokyo | Fluorescent probe |
| CN103342697A (en) * | 2013-07-05 | 2013-10-09 | 中国科学院合肥物质科学研究院 | Difunctional near-infrared fluorescence molecular probe for detecting hypochlorous acid and preparation method for same |
| CN103555321A (en) * | 2013-10-30 | 2014-02-05 | 南京邮电大学 | Phosphorescent ionic type iridium complex probe and preparation method and application thereof |
| CN108102645A (en) * | 2017-12-12 | 2018-06-01 | 南京邮电大学 | The preparation and application of a kind of fluorescence probe for hypochlorite ion's detection |
| CN108218923A (en) * | 2018-03-06 | 2018-06-29 | 南京邮电大学 | A kind of water-soluble phosphorus photopolymer for detecting pH and its preparation method and application |
| CN108535233A (en) * | 2018-05-17 | 2018-09-14 | 南京邮电大学 | One kind is for hypochlorous water-soluble phosphorescence nano-particle of ratio method detection and the preparation method and application thereof |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3483610A1 (en) * | 2009-11-09 | 2019-05-15 | University Of Washington Center For Commercialization | Functionalized chromophoric polymer dots and bioconjugates thereof |
| CN102206235A (en) * | 2011-03-18 | 2011-10-05 | 南京邮电大学 | Phosphorescent iridium complex capable of dissolving in water system, and preparation method thereof |
| US9788765B2 (en) * | 2012-09-28 | 2017-10-17 | Dexcom, Inc. | Zwitterion surface modifications for continuous sensors |
| WO2016015027A1 (en) * | 2014-07-25 | 2016-01-28 | SeLux Diagnostics, Inc. | Assay methods involving dissociable nanoparticles |
| CN106046059B (en) * | 2016-06-06 | 2018-08-17 | 南京邮电大学 | A kind of phosphorescent iridium complex probe and its preparation and application with Mitochondrially targeted function |
| CN106281308A (en) * | 2016-07-29 | 2017-01-04 | 南京邮电大学 | A kind of polymer nano-particle being coated with ionic type iridium complex and its preparation method and application |
| CN106892947B (en) * | 2017-02-28 | 2019-07-30 | 陕西师范大学 | One kind containing the complex of iridium and its preparation method and application of (Hydrazinocarbonyl) ferrocene ligands |
-
2018
- 2018-05-17 CN CN201810473705.3A patent/CN108535233B/en active Active
- 2018-10-24 WO PCT/CN2018/111540 patent/WO2019218594A1/en active Application Filing
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007100061A1 (en) * | 2006-03-03 | 2007-09-07 | The University Of Tokyo | Fluorescent probe |
| CN103342697A (en) * | 2013-07-05 | 2013-10-09 | 中国科学院合肥物质科学研究院 | Difunctional near-infrared fluorescence molecular probe for detecting hypochlorous acid and preparation method for same |
| CN103555321A (en) * | 2013-10-30 | 2014-02-05 | 南京邮电大学 | Phosphorescent ionic type iridium complex probe and preparation method and application thereof |
| CN108102645A (en) * | 2017-12-12 | 2018-06-01 | 南京邮电大学 | The preparation and application of a kind of fluorescence probe for hypochlorite ion's detection |
| CN108218923A (en) * | 2018-03-06 | 2018-06-29 | 南京邮电大学 | A kind of water-soluble phosphorus photopolymer for detecting pH and its preparation method and application |
| CN108535233A (en) * | 2018-05-17 | 2018-09-14 | 南京邮电大学 | One kind is for hypochlorous water-soluble phosphorescence nano-particle of ratio method detection and the preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108535233A (en) | 2018-09-14 |
| CN108535233B (en) | 2020-07-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | A Hydrogen‐Bonded‐Supramolecular‐Polymer‐Based Nanoprobe for Ratiometric Oxygen Sensing in Living Cells | |
| WO2019218594A1 (en) | Water-soluble phosphorescent nanoparticle for detecting hypochlorous acid using ratio method and preparation method and application thereof | |
| Su et al. | AIE-based Tb3+ complex self-assembled nanoprobe for ratiometric fluorescence detection of anthrax spore biomarker in water solution and actual spore samples | |
| CN110128414B (en) | Preparation and application of hemicyanine dye-based hypoxia fluorescent probe | |
| CN106084873B (en) | A kind of efficiently near-infrared fluorescent material and its biologic applications | |
| CN113135906A (en) | Lipid drop targeted fluorescent probe capable of specifically detecting polarity change in lipid drop | |
| CN115386371B (en) | Blue fluorescent quantum dot preparation method and application of blue fluorescent quantum dot as glutathione detection probe | |
| Zhang et al. | Selective visualization of endogenous hydrogen sulfide in lysosomes using aggregation induced emission dots | |
| KR20110120224A (en) | Water soluble fluorescent fullerene derivatives and method for preparing the same | |
| Zhao et al. | Self-enhanced carbonized polymer dots for selective visualization of lysosomes and real-time apoptosis monitoring | |
| Khan et al. | Solvent-free synthesis of nitrogen doped carbon dots with dual emission and their biological and sensing applications | |
| US20070092927A1 (en) | Photocatalytic particles with directed and controlled redox activity | |
| CN113201335B (en) | Enhanced fluorescent carbon dot, preparation method and application in cadmium ion detection | |
| CN113604213B (en) | Nano fluorescent probe and preparation method and application thereof | |
| Liu et al. | Hydrothermal synthesis of a highly photoluminescent molecule from citric acid and cysteamine for the efficient detection of Au3+ in aqueous solution | |
| Zhang et al. | In vivo selective imaging of metabolic glycosylation with a tetrazine-modified upconversion nanoprobe | |
| CN110372681B (en) | Application of self-assembled nano fluorescent probe for selectively detecting human serum albumin | |
| CN110669503B (en) | Preparation and application of carbon monoxide near-infrared fluorescent probe | |
| Sun et al. | One-pot synthesis of conjugated polymer dots with ultrasmall size below 10 nm through Schiff-base chemistry and their bioapplications in monitoring lysosomal pH | |
| Gao et al. | One-pot synthesis of efficient multifunctional nitrogen-doped carbon dots with efficient yellow fluorescence emission for detection of hypochlorite and thiosulfate | |
| Shen et al. | A one-step synthesis of novel high pH-sensitive nitrogen-doped yellow fluorescent carbon dots and their detection application in living cells | |
| CN110117236A (en) | It is a kind of using TS as small-molecule fluorescent probe of fluorogen and the preparation method and application thereof | |
| CN113024504B (en) | Preparation and application of hydrogen sulfide fluorescent probe based on isophorone-xanthene | |
| Li et al. | A platinum (II)-acetylide-based conjugated polyelectrolyte for hypoxia imaging via ratiometric and time-resolved luminescence microscopy | |
| CN112239462B (en) | Cyanide receptor compound based on coumarin and carboxylic indole, preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 18918818 Country of ref document: EP Kind code of ref document: A1 |
|
| NENP | Non-entry into the national phase |
Ref country code: DE |
|
| 122 | Ep: pct application non-entry in european phase |
Ref document number: 18918818 Country of ref document: EP Kind code of ref document: A1 |