CN108752377A - 一种检测过氧亚硝基阴离子的荧光探针、合成方法和应用 - Google Patents
一种检测过氧亚硝基阴离子的荧光探针、合成方法和应用 Download PDFInfo
- Publication number
- CN108752377A CN108752377A CN201810684964.0A CN201810684964A CN108752377A CN 108752377 A CN108752377 A CN 108752377A CN 201810684964 A CN201810684964 A CN 201810684964A CN 108752377 A CN108752377 A CN 108752377A
- Authority
- CN
- China
- Prior art keywords
- fluorescence probe
- detection
- peroxynitrite
- synthetic method
- onoo
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000000523 sample Substances 0.000 title claims abstract description 70
- 238000001514 detection method Methods 0.000 title claims abstract description 34
- 238000010189 synthetic method Methods 0.000 title claims abstract description 11
- CMFNMSMUKZHDEY-UHFFFAOYSA-N peroxynitrous acid Chemical compound OON=O CMFNMSMUKZHDEY-UHFFFAOYSA-N 0.000 title abstract 3
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims abstract description 18
- HUMNYLRZRPPJDN-UHFFFAOYSA-N benzaldehyde Chemical compound O=CC1=CC=CC=C1 HUMNYLRZRPPJDN-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000013067 intermediate product Substances 0.000 claims abstract description 9
- 238000012545 processing Methods 0.000 claims abstract description 9
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims abstract description 6
- QNGNSVIICDLXHT-UHFFFAOYSA-N para-ethylbenzaldehyde Natural products CCC1=CC=C(C=O)C=C1 QNGNSVIICDLXHT-UHFFFAOYSA-N 0.000 claims abstract description 6
- QPQGTZMAQRXCJW-UHFFFAOYSA-N [chloro(phenyl)phosphoryl]benzene Chemical compound C=1C=CC=CC=1P(=O)(Cl)C1=CC=CC=C1 QPQGTZMAQRXCJW-UHFFFAOYSA-N 0.000 claims abstract description 5
- NWZSZGALRFJKBT-KNIFDHDWSA-N (2s)-2,6-diaminohexanoic acid;(2s)-2-hydroxybutanedioic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O.NCCCC[C@H](N)C(O)=O NWZSZGALRFJKBT-KNIFDHDWSA-N 0.000 claims abstract description 4
- IKDUDTNKRLTJSI-UHFFFAOYSA-N hydrazine monohydrate Substances O.NN IKDUDTNKRLTJSI-UHFFFAOYSA-N 0.000 claims abstract description 4
- 239000003960 organic solvent Substances 0.000 claims abstract description 4
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical group NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 claims abstract description 3
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims abstract description 3
- CMFNMSMUKZHDEY-UHFFFAOYSA-M peroxynitrite Chemical compound [O-]ON=O CMFNMSMUKZHDEY-UHFFFAOYSA-M 0.000 claims description 14
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 12
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 8
- -1 nitroso anion Chemical class 0.000 claims description 7
- 238000010898 silica gel chromatography Methods 0.000 claims description 6
- 150000002978 peroxides Chemical class 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Natural products CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 3
- 239000003208 petroleum Substances 0.000 claims description 3
- 235000019441 ethanol Nutrition 0.000 claims description 2
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 claims description 2
- 229910052760 oxygen Inorganic materials 0.000 claims description 2
- 239000001301 oxygen Substances 0.000 claims description 2
- 125000005909 ethyl alcohol group Chemical group 0.000 claims 1
- 238000006243 chemical reaction Methods 0.000 abstract description 15
- 238000006073 displacement reaction Methods 0.000 abstract description 10
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 28
- 239000000243 solution Substances 0.000 description 16
- 238000000034 method Methods 0.000 description 8
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 230000003647 oxidation Effects 0.000 description 4
- 238000007254 oxidation reaction Methods 0.000 description 4
- 238000003756 stirring Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000002189 fluorescence spectrum Methods 0.000 description 3
- 239000012074 organic phase Substances 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- AVXURJPOCDRRFD-UHFFFAOYSA-N Hydroxylamine Chemical compound ON AVXURJPOCDRRFD-UHFFFAOYSA-N 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 238000013019 agitation Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000006701 autoxidation reaction Methods 0.000 description 2
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 239000012043 crude product Substances 0.000 description 2
- 238000000695 excitation spectrum Methods 0.000 description 2
- 238000002284 excitation--emission spectrum Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 230000003760 hair shine Effects 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 238000004020 luminiscence type Methods 0.000 description 2
- NUJOXMJBOLGQSY-UHFFFAOYSA-N manganese dioxide Inorganic materials O=[Mn]=O NUJOXMJBOLGQSY-UHFFFAOYSA-N 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000010791 quenching Methods 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000002371 ultraviolet--visible spectrum Methods 0.000 description 2
- 238000009834 vaporization Methods 0.000 description 2
- 230000008016 vaporization Effects 0.000 description 2
- AXKGIPZJYUNAIW-UHFFFAOYSA-N (4-aminophenyl)methanol Chemical class NC1=CC=C(CO)C=C1 AXKGIPZJYUNAIW-UHFFFAOYSA-N 0.000 description 1
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- NJLHHACGWKAWKL-UHFFFAOYSA-N ClP(Cl)=O Chemical compound ClP(Cl)=O NJLHHACGWKAWKL-UHFFFAOYSA-N 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- JCXJVPUVTGWSNB-UHFFFAOYSA-N Nitrogen dioxide Chemical compound O=[N]=O JCXJVPUVTGWSNB-UHFFFAOYSA-N 0.000 description 1
- ODUCDPQEXGNKDN-UHFFFAOYSA-N Nitrogen oxide(NO) Natural products O=N ODUCDPQEXGNKDN-UHFFFAOYSA-N 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- 206010057430 Retinal injury Diseases 0.000 description 1
- 208000034189 Sclerosis Diseases 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 239000004305 biphenyl Substances 0.000 description 1
- 235000010290 biphenyl Nutrition 0.000 description 1
- 125000006267 biphenyl group Chemical group 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000005281 excited state Effects 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 150000002240 furans Chemical class 0.000 description 1
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000000802 nitrating effect Effects 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 150000004893 oxazines Chemical class 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- ZUOUZKKEUPVFJK-UHFFFAOYSA-N phenylbenzene Natural products C1=CC=CC=C1C1=CC=CC=C1 ZUOUZKKEUPVFJK-UHFFFAOYSA-N 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 238000006862 quantum yield reaction Methods 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229940065287 selenium compound Drugs 0.000 description 1
- 150000003343 selenium compounds Chemical class 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
Classifications
-
- 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
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/28—Phosphorus compounds with one or more P—C bonds
- C07F9/30—Phosphinic acids [R2P(=O)(OH)]; Thiophosphinic acids ; [R2P(=X1)(X2H) (X1, X2 are each independently O, S or Se)]
- C07F9/32—Esters thereof
- C07F9/3205—Esters thereof the acid moiety containing a substituent or a structure which is considered as characteristic
- C07F9/3229—Esters of aromatic acids (P-C aromatic linkage)
-
- 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
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/28—Phosphorus compounds with one or more P—C bonds
- C07F9/30—Phosphinic acids [R2P(=O)(OH)]; Thiophosphinic acids ; [R2P(=X1)(X2H) (X1, X2 are each independently O, S or Se)]
- C07F9/32—Esters thereof
- C07F9/3258—Esters thereof the ester moiety containing a substituent or a structure which is considered as characteristic
- C07F9/3282—Esters with hydroxyaryl 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
-
- 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"
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1007—Non-condensed systems
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1003—Carbocyclic compounds
- C09K2211/1014—Carbocyclic compounds bridged by heteroatoms, e.g. N, P, Si or B
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Materials Engineering (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Optics & Photonics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
本发明公开了一种检测过氧亚硝基阴离子的荧光探针、合成方法和应用,所述荧光探针的结构式为:所述荧光探针的Stoke位移为120nm,吸收波长的范围为450‑476nm,应用于对过氧亚硝基阴离子进行定量检测,其合成方法包括将2‑羟基苯甲醛和三乙胺溶于四氢呋喃中,加入二苯基次膦酰氯与之反应;反应完成后,处理得到中间产物;将中间产物与水合肼溶于有机溶剂中,处理得到荧光探针。本发明的荧光探针选择性高,在检测ONOO‑时能产生较大的Stoke位移,能提高检测ONOO‑的精准度。
Description
技术领域
本发明涉及应用生物领域,具体涉及到一种检测过氧亚硝基阴离子的荧光探针、合成方法和应用。
背景技术
过氧亚硝基阴离子(ONOO-)被认为是最重要的阴离子之一,在人体内由过量的一氧化氮(NO)和氧自由基(.O2-)反应产生,它既是强氧化剂,又是硝化剂,可与人体细胞内的核酸、蛋白质、脂质等大分子物质反应,使细胞的代谢发生困难并且使机体消耗大量的能量,从而引起一系列的生物病变过程,如肿瘤、动脉硬化、身体炎症、视网膜损伤等。近年来,医学界也不乏大量对于ONOO-的检测的研究,但是由于ONOO-的强氧化性,在体内很不稳定,这为ONOO-的检测带来不便。因此,建立一种快速、灵敏、高选择性的检测方法用于研究过氧亚硝基阴离子引起的生物体病变具有十分重要的医学意义。目前,检测ONOO-的方法主要有电子自旋法,紫外光谱法,电分析法以及化学发光法,但是这些方法大部分比较复杂,耗时、耗力、干扰因素多,有的还需要贵重仪器和较大的样品体积,一般不适用于生物体内快速、实时、原位定量对ONOO-进行检测和分析。因此,研究一种新的方法检测生物体内ONOO-具有重要的意义。
荧光探针法具有高选择性和灵敏度,非破坏性的分析和检测***易操作性等优点,在环境、化学、生物和医学等领域存在着广泛的应用。其中,以有机荧光染料为基础的生物分子标记和分子荧光探针具有操作简便、重现性好等优点,可方便用于生物分子的原位、实时无损伤检测以及生物分子及其生物过程的追踪。然而,大多数荧光染料(如荧光素,罗丹明,恶嗪和花菁素)选择性不高,并不能区分开ONOO-同其他活性氧化物,且产生的Stokes位移都非常小(一般<30nm),导致激发光谱和发射光谱之间的严重串扰,造成成像时信噪比低和严重的荧光自淬灭现象,限制了其在生物成像中的应用。
目前,已报道的检测ONOO-的荧光机制主要为机硒化合物的氧化、4-羟甲基苯胺化合物氧化、肼类化合物的氧化等。然而,对生物体内ONOO-检测,这些荧光物质不仅Stoke位移较小,还容易与其他氧化物如·OH、OCl-发生反应;特别是较小的Stoke位移,比较容易受到生物体自身的干扰,不利于ONOO-的精准检测。
文献CN107488189A公开了二酚化合物与2-醛基苯硼酸形成的自组装及其作为荧光探针的应用,用于对自由基次氯酸和过氧亚硝基的定量检测,但该方法并没有涉及和解决荧光探针stoke位移较小和选择性不高的问题。
发明内容
本发明要解决的技术问题是提供一种检测过氧亚硝基阴离子的荧光探针、合成方法和应用,该荧光探针选择性高,在检测ONOO-时能产生较大的Stoke位移,能提高检测ONOO-的精准度。
本发明的内容包括荧光探针的结构如式Ⅰ所示:
其中,R基团能替换为-OH、-Cl和-OCH3,荧光探针的Stoke能位移达到120nm。
所述荧光探针吸收波长的范围为450-476nm。
荧光探针本身没有荧光,但经过氧亚硝基氧化后生成有较强荧光的荧光团,通过观测荧光团的荧光强度的变化来达到定量检测的目的。
本发明的荧光探针加入ONOO-后,反应控制在pH=7.4(其中荧光探针与ONOO-作用前后荧光强度随pH的变化如图7所示),温度为37℃的条件下,产生的荧光团结构如式Ⅱ所示:
其中,反应后荧光团的产量为52-74%,氧化后荧光团的量子产量为12%,该荧光团具有聚集诱导荧光发射(AIE)和激发态分子内质子转移(ESIPT)机理的性质,因此这种荧光染料能产生较大的Stoke位移。
将荧光探针与其他活性氧化物(如·OH、·O2-、NO)分别进行反应,发现只有过氧亚硝基可以与荧光探针反应产生如式Ⅱ所示的荧光团,验证了荧光探针的高选择性。
还包括荧光探针的以下合成步骤:
(1)将2-羟基苯甲醛和三乙胺溶于干燥的四氢呋喃中,搅拌下逐滴加入二苯基次膦酰氯,室温下反应12小时,其中2-羟基苯甲醛、三乙胺和二苯基次膦酰氯的摩尔比为1:2:1;
(2)反应完成后,减压蒸发得到粗产品,然后用硅胶柱层析处理得到中间产物,在硅胶柱层析处理中乙酸乙酯和石油醚的体积比为1:4;
(3)将中间产物与水合肼溶于有机溶剂中,将反应混合物经过加热、搅拌、回流处理,处理时间为2小时,待反应液冷却后,抽取滤液,得到荧光探针。
所述荧光探针用于对过氧亚硝基阴离子进行定量检测。
本发明的有益效果是,该荧光探针合成简单、响应快,并且选择性高,在检测ONOO-时具有较大的Stoke位移,能更加精准的检测ONOO-。
附图说明
图1为中间产物的合成反应方程式。
图2为荧光探针的合成反应方程式。
图3为荧光探针的1HNMR图谱。
图4为荧光探针的13C NMR图谱。
图5为荧光探针加入(蓝色)和不加(红色)ONOO-的紫外可见光吸收光谱图。
图6为荧光探针与ONOO-作用后的荧光强度图。
图7为荧光探针与ONOO-作用前后荧光强度随pH的变化图。
图8为荧光探针Stoke位移光谱图。
图9为荧光探针与不同分析物作用的荧光强度变化图。
图10为荧光探针与ONOO-作用前后荧光强度随时间的变化图。
具体实施方式
实施例1:
关于荧光探针的合成方法,包括以下实验步骤:
(1)将2-羟基苯甲醛(122mg,1mmol)和三乙胺(200mg,2mmol)溶于干燥的10mL四氢呋喃中,搅拌下逐滴加入二苯基次膦酰氯(236.5mg,1mmol),室温下反应12小时;
(2)反应完成后,减压蒸发得到粗产品,然后用硅胶柱层析处理,其中硅胶柱层析处理中乙酸乙酯和石油醚的体积比为1:4,得到白色中间产物(232mg,产率72%),该中间产物的合成反应方程式如图1所示;
(3)将白色中间产物(322mg,1mmol)与水合肼(50mg,1mmol)溶于10mL乙醇中,将反应混合物经过加热、搅拌、回流处理,处理时间为2小时,待反应液冷却后,抽取滤液,得到黄色固体(288mg,产率45%),该黄色固体—即荧光探针的合成反应方程式如图2所示。
所述荧光探针的核磁共振谱为:
1H NMR(500MHz,CDCl3)δ(ppm):8.96(s,2H),8.10(d,J=7,2H),7.92(dd,J=7.5Hz,J=12.5Hz,18H),7.54(m,6H),7.47(m,8H),7.34(t,J=8,2H),7.18(t,J=7.5Hz,2H);其中,1H NMR图谱如图3所示;
13C NMR(125MHz,CDCl3)δ(ppm):157.1,151.0,132.7,132.7,132.4,131.8,131.7,131.0,129.9,128.8,128.7,128.2,125.2,124.7,121.1;13C NMR图谱如图4所示。
实施例2:
关于检验荧光探针检测ONOO-的效果,其实验步骤为:
(1)将荧光探针溶于N,N-二甲基甲酰胺(DMF)中,配置成1mmol/L的探针溶液;
(2)取探针溶液加入DMF和PBS(pH=7.4)缓冲液,配置成10μM(有机相:PBS水相=1:99,V/V)的溶液,测试紫外吸收光谱和荧光发射光谱变化情况。
紫外吸收光谱的变化情况如图5所示,其中荧光探针加入ONOO-的紫外可见光吸收光谱曲线为(a),不加ONOO-的紫外可见光吸收光谱曲线为(b)。从图5中可以看出,当不加入ONOO-的情况下,探针在400nm处没有吸收峰,而加入ONOO-之后,探针在400nm处出现了明显的吸收峰,说明本荧光探针对ONOO-具有检测效果。
荧光发射光谱变化情况如图6所示,从图6可以看出,荧光发射光谱在Em=520nm处随着ONOO-浓度的增加荧光逐渐增强,其中ONOO-浓度变化为0,5,10,15,20,25,30,35,40,45uM。
实施例3:
为检验本发明荧光探针的Stoke位移值,现做以下实验。
(1)通过羟胺在碱性介质中自氧化合成ONOO-:在有氧条件下激烈搅拌含0.01mol/L羟胺、0.5mol/LNaOH和0.001mol/L EDTA的混合液约3小时,然后用MnO2粉末过滤混合液除去H2O2,过滤后的混合液可立即用于实验,也可于-18℃下贮存。其中,可在紫外可见分光光度计302nm处检测ONOO-浓度。
(2)将荧光探针溶于N,N-二甲基甲酰胺(DMF)中,配置成1mmol/L的探针溶液,取探针溶液加入DMF和PBS(pH=7.4)缓冲液,配置成10uM(有机相:PBS水相=1:99,V/V)的溶液;再加入45uM ONOO-反应液,延迟10秒,连续测其10秒发光强度,取发光峰值,计算发光抑制率。
如图8所示,本发明的荧光探针与ONOO-反应后,产生了较大的Stoke位移,Stoke位移达到了120nm左右,具有较高的区分度,能减少激发光谱和发射光谱之间的串扰和减弱荧光自淬灭现象,最终提高ONOO-检测的精准度。
实施例4:
为检验本发明荧光探针在ONOO-选择性上专一反应的表现,现做以下实验。
包括以下实验步骤:
(1)将荧光探针溶于N,N-二甲基甲酰胺(DMF)中,配置成1mmol/L的探针溶液;
(2)取探针溶液加入DMF和PBS(pH=7.4)缓冲液,再分别加入45uM的-O2,t-BuOO-,H2O2,ClO-,·OH,NO,NO3 -,HNO,NO2 -,Fe3+,Cu2+,Zn2+,Cr3+反应液与之反应;
(3)延迟10秒,连续测其10秒发光强度,取发光峰值,计算发光抑制率。
从图9中可以看出,荧光探针只对ONOO-具有较高的选择性,基本不与其他活性氧化物发生反应。
实施例5:
为了测试时间对荧光强度的影响,对探针化合物与ONOO-作用前后的荧光强度随时间的变化进行了相关实验。
实验步骤为:
(1)通过羟胺在碱性介质中自氧化合成ONOO-:在有氧条件下激烈搅拌含0.01mol/L羟胺、0.5mol/LNaOH和0.001mol/LEDTA的混合液约3小时,然后用MnO2粉末过滤混合液除去H2O2,过滤后的混合液可立即用于实验,也可于-18℃下贮存。其中,可在紫外可见分光光度计302nm处检测ONOO-浓度。
(2)将荧光探针溶于N,N-二甲基甲酰胺(DMF)中,配置成1mmol/L的探针溶液,取探针溶液加入DMF和PBS(pH=7.4)缓冲液,配置成10μM(有机相:PBS水相=1:99,V/V)的溶液;再加入45uM ONOO-反应液,延迟10秒,每隔10秒测其发光强度。
从图10可以看出,探针化合物与ONOO-作用后,随着时间的变化荧光强度逐渐增强,在18分钟左右荧光达到最大值,说明探针化合物能够较快的检测ONOO-。
Claims (10)
1.一种检测过氧亚硝基阴离子的荧光探针,其特征在于,其结构如式Ⅰ所示:
其中,R基团为-H、-OH、-Cl或-OCH3。
2.一种如权利要求1所述的检测过氧亚硝基阴离子的荧光探针的合成方法,其特征在于,包括以下步骤:
(1)将2-羟基苯甲醛和三乙胺溶于四氢呋喃中,加入二苯基次膦酰氯与之反应;
(2)反应完成后,处理得到中间产物;
(3)将中间产物与水合肼溶于有机溶剂中,处理得到荧光探针。
3.如权利要求2所述的合成方法,其特征在于:所述步骤(1)中,2-羟基苯甲醛、三乙胺和二苯基次膦酰氯的摩尔比为1:2:1。
4.如权利要求2所述的合成方法,其特征在于:所述步骤(2)中的处理包括减压蒸发并用硅胶柱层析。
5.如权利要求4所述的合成方法,其特征在于:所述硅胶柱层析中使用乙酸乙酯和石油醚进行洗脱,所述乙酸乙酯和石油醚的体积比为1:4。
6.如权利要求2所述的合成方法,其特征在于:所述步骤(3)中,有机溶剂为乙醇。
7.一种如权利要求1所述的检测过氧亚硝基阴离子的荧光探针的应用,其特征在于:所述荧光探针用于对过氧亚硝基阴离子进行定量检测。
8.如权利要求7所述的检测过氧亚硝基阴离子的荧光探针的应用,其特征在于:对过氧亚硝基阴离子进行定量检测时,荧光探针吸收波长的范围为450-476nm。
9.如权利要求7所述的检测过氧亚硝基阴离子的荧光探针的应用,其特征在于:对过氧亚硝基阴离子进行定量检测时,pH值为7.4。
10.如权利要求7所述的检测过氧亚硝基阴离子的荧光探针的应用,其特征在于:对过氧亚硝基阴离子进行定量检测时,温度控制在37℃。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810684964.0A CN108752377B (zh) | 2018-06-28 | 2018-06-28 | 一种检测过氧亚硝基阴离子的荧光探针、合成方法和应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201810684964.0A CN108752377B (zh) | 2018-06-28 | 2018-06-28 | 一种检测过氧亚硝基阴离子的荧光探针、合成方法和应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN108752377A true CN108752377A (zh) | 2018-11-06 |
CN108752377B CN108752377B (zh) | 2020-07-03 |
Family
ID=63974207
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201810684964.0A Active CN108752377B (zh) | 2018-06-28 | 2018-06-28 | 一种检测过氧亚硝基阴离子的荧光探针、合成方法和应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN108752377B (zh) |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110669501A (zh) * | 2019-10-25 | 2020-01-10 | 南京林业大学 | 一种对过氧亚硝基阴离子响应的近红外荧光探针及其制备方法和应用 |
CN111285833A (zh) * | 2020-02-12 | 2020-06-16 | 曲阜师范大学 | 一种检测onoo-的比率型荧光分子探针及其制备方法和应用 |
CN112574252A (zh) * | 2020-11-24 | 2021-03-30 | 河南大学 | 基于试卤灵染料专一响应onoo-的荧光探针、制备方法及应用 |
CN115850314A (zh) * | 2022-12-30 | 2023-03-28 | 厦门大学附属心血管病医院 | 一种过氧亚硝酸盐和脂滴双响应探针及其制备方法、应用 |
CN117756835A (zh) * | 2024-02-22 | 2024-03-26 | 烟台大学 | 一种近红外检测过氧亚硝基的荧光探针及其制备方法和应用 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1837211A (zh) * | 2006-04-11 | 2006-09-27 | 山东师范大学 | 一种检测超氧阴离子自由基的荧光探针及其合成方法和用途 |
CN107488189A (zh) * | 2017-08-14 | 2017-12-19 | 西北农林科技大学 | 二酚化合物与2‑醛基苯硼酸形成的自组装及其作为荧光探针的应用 |
-
2018
- 2018-06-28 CN CN201810684964.0A patent/CN108752377B/zh active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1837211A (zh) * | 2006-04-11 | 2006-09-27 | 山东师范大学 | 一种检测超氧阴离子自由基的荧光探针及其合成方法和用途 |
CN107488189A (zh) * | 2017-08-14 | 2017-12-19 | 西北农林科技大学 | 二酚化合物与2‑醛基苯硼酸形成的自组装及其作为荧光探针的应用 |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110669501A (zh) * | 2019-10-25 | 2020-01-10 | 南京林业大学 | 一种对过氧亚硝基阴离子响应的近红外荧光探针及其制备方法和应用 |
CN110669501B (zh) * | 2019-10-25 | 2023-03-03 | 南京林业大学 | 一种对过氧亚硝基阴离子响应的近红外荧光探针及其制备方法和应用 |
CN111285833A (zh) * | 2020-02-12 | 2020-06-16 | 曲阜师范大学 | 一种检测onoo-的比率型荧光分子探针及其制备方法和应用 |
CN112574252A (zh) * | 2020-11-24 | 2021-03-30 | 河南大学 | 基于试卤灵染料专一响应onoo-的荧光探针、制备方法及应用 |
CN115850314A (zh) * | 2022-12-30 | 2023-03-28 | 厦门大学附属心血管病医院 | 一种过氧亚硝酸盐和脂滴双响应探针及其制备方法、应用 |
CN115850314B (zh) * | 2022-12-30 | 2024-06-21 | 厦门大学附属心血管病医院 | 一种过氧亚硝酸盐和脂滴双响应探针及其制备方法、应用 |
CN117756835A (zh) * | 2024-02-22 | 2024-03-26 | 烟台大学 | 一种近红外检测过氧亚硝基的荧光探针及其制备方法和应用 |
CN117756835B (zh) * | 2024-02-22 | 2024-04-30 | 烟台大学 | 一种近红外检测过氧亚硝基的荧光探针及其制备方法和应用 |
Also Published As
Publication number | Publication date |
---|---|
CN108752377B (zh) | 2020-07-03 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN108752377A (zh) | 一种检测过氧亚硝基阴离子的荧光探针、合成方法和应用 | |
Gunnlaugsson et al. | Novel sodium-selective fluorescent PET and optically based chemosensors: towards Na+ determination in serum | |
Su et al. | A near-infrared and colorimetric fluorescent probe for palladium detection and bioimaging | |
CN106281304B (zh) | 一种可用于活细胞内丙二醛成像的荧光探针及其制备方法 | |
CN104017568B (zh) | 一种含罗丹明的荧光探针在检测Hg2+上的应用 | |
Hu et al. | A ratiometric fluorescence sensor for Fe3+ based on FRET and PET processes | |
CN106432348A (zh) | 一种基于铕配合物的可见光可激发的比率荧光温敏探针及其制备方法和应用 | |
Pan et al. | AIE fluorescent probe based on tetraphenylethylene and morpholine-thiourea structures for detection of HClO | |
Du et al. | A new coumarin-based “turn-on” fluorescence probe with high sensitivity and specificity for detecting hypochlorite ion | |
CN104830312B (zh) | 一种荧光增强型探针化合物的制备以及三价铬离子检测 | |
Huang et al. | A pyridyl functionalized rhodamine chemodosimeter for selective fluorescent detection of mercury ions and cell imaging | |
CN109021000B (zh) | 一种检测过氧化氢的荧光探针、合成方法和应用 | |
CN113861076A (zh) | 一种aie型三苯胺衍生物荧光探针及其制备方法和其在水合肼检测中的应用 | |
CN111848671B (zh) | 一种过氧亚硝酰阴离子比率远红外荧光探针、制备方法和应用 | |
Ye et al. | Design and synthesis of a new terbium complex-based luminescent probe for time-resolved luminescence sensing of zinc ions | |
CN105968098A (zh) | 一种含咔唑、苯并咪唑取代的喹啉衍生物及其制备方法和应用 | |
CN110357896A (zh) | 一类化合物及制备与其在检测二价铜离子和强酸pH中的应用 | |
Hashemi et al. | Pyridine-2-yl Quinoxaline (2-CPQ) Derivative As a Novel Pink Fluorophore: Synthesis, and Chemiluminescence Characteristics | |
CN110590784B (zh) | 一种基于吡咯并吡咯二酮的衍生物及制备方法与应用 | |
CN109721592B (zh) | 一种含香豆素的氨基吡嗪酰腙衍生物的荧光探针及其制备方法和应用 | |
CN105884710B (zh) | 一种基于杯芳烃的1,3,4-噁二唑类Cu2+荧光探针及其合成方法 | |
CN110467570A (zh) | 一种四苯乙烯-8-羟基喹啉类化合物及其制备方法和应用 | |
CN111187207B (zh) | 一种双酰胺基喹啉类探针分子及其制备方法和应用 | |
CN109705055A (zh) | 苯并噻唑类化合物及其制备方法及作为次溴酸探针的应用 | |
Cao et al. | A ratiometric fluorescent probe for hydrazine imaging in biological and environmental samples |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant | ||
TR01 | Transfer of patent right |
Effective date of registration: 20230329 Address after: No. 10 Changgang Road, Jianggao Town, Baiyun District, Guangzhou City, Guangdong Province, 510470 Patentee after: Yuewang Agricultural Group Co.,Ltd. Address before: 415000 3150 Dongting Road, Wuling District, Changde, Hunan Patentee before: HUNAN University OF ARTS AND SCIENCE |
|
TR01 | Transfer of patent right |