CN111848671B - 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 - Google Patents
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 Download PDFInfo
- Publication number
- CN111848671B CN111848671B CN202010745792.0A CN202010745792A CN111848671B CN 111848671 B CN111848671 B CN 111848671B CN 202010745792 A CN202010745792 A CN 202010745792A CN 111848671 B CN111848671 B CN 111848671B
- Authority
- CN
- China
- Prior art keywords
- probe
- ani
- fluorescent probe
- far infrared
- 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.)
- Active
Links
- 239000007850 fluorescent dye Substances 0.000 title claims abstract description 30
- -1 peroxide anion Chemical class 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title abstract description 7
- 238000003745 diagnosis Methods 0.000 claims abstract 2
- 150000001875 compounds Chemical class 0.000 claims description 26
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 7
- 150000001450 anions Chemical class 0.000 claims description 5
- HVTICUPFWKNHNG-UHFFFAOYSA-N iodoethane Chemical compound CCI HVTICUPFWKNHNG-UHFFFAOYSA-N 0.000 claims description 5
- 239000003960 organic solvent Substances 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 4
- 238000010992 reflux Methods 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 2
- 201000010099 disease Diseases 0.000 claims 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims 1
- 238000001514 detection method Methods 0.000 abstract description 14
- 238000006073 displacement reaction Methods 0.000 abstract description 4
- 230000003902 lesion Effects 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 44
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 9
- 230000008859 change Effects 0.000 description 9
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 8
- 239000000243 solution Substances 0.000 description 6
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
- BHIIGRBMZRSDRI-UHFFFAOYSA-N [chloro(phenoxy)phosphoryl]oxybenzene Chemical compound C=1C=CC=CC=1OP(=O)(Cl)OC1=CC=CC=C1 BHIIGRBMZRSDRI-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- IOJUPLGTWVMSFF-UHFFFAOYSA-N benzothiazole Chemical class C1=CC=C2SC=NC2=C1 IOJUPLGTWVMSFF-UHFFFAOYSA-N 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N coumarin Chemical compound C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000002189 fluorescence spectrum Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 2
- 238000004435 EPR spectroscopy Methods 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- 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 description 1
- 238000004847 absorption spectroscopy Methods 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 235000010290 biphenyl Nutrition 0.000 description 1
- 239000004305 biphenyl Substances 0.000 description 1
- 239000012496 blank sample Substances 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 229960000956 coumarin Drugs 0.000 description 1
- 235000001671 coumarin Nutrition 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000002848 electrochemical method Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- QWPPOHNGKGFGJK-UHFFFAOYSA-N hypochlorous acid Chemical compound ClO QWPPOHNGKGFGJK-UHFFFAOYSA-N 0.000 description 1
- 238000012308 immunohistochemistry method Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 238000005935 nucleophilic addition reaction Methods 0.000 description 1
- 230000000269 nucleophilic effect Effects 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- ZUOUZKKEUPVFJK-UHFFFAOYSA-N phenylbenzene Natural products C1=CC=CC=C1C1=CC=CC=C1 ZUOUZKKEUPVFJK-UHFFFAOYSA-N 0.000 description 1
- 239000002953 phosphate buffered saline Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 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
- INCIMLINXXICKS-UHFFFAOYSA-M pyronin Y Chemical compound [Cl-].C1=CC(=[N+](C)C)C=C2OC3=CC(N(C)C)=CC=C3C=C21 INCIMLINXXICKS-UHFFFAOYSA-M 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 239000011669 selenium Substances 0.000 description 1
- 238000011896 sensitive detection Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- ANRHNWWPFJCPAZ-UHFFFAOYSA-M thionine Chemical compound [Cl-].C1=CC(N)=CC2=[S+]C3=CC(N)=CC=C3N=C21 ANRHNWWPFJCPAZ-UHFFFAOYSA-M 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 238000002211 ultraviolet spectrum Methods 0.000 description 1
Images
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
- C07F9/3258—Esters thereof the ester moiety containing a substituent or a structure which is considered as characteristic
-
- 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
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N2021/6417—Spectrofluorimetric devices
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Optics & Photonics (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
本发明涉及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，过氧亚硝酰阴离子比率远红外荧光探针的结构式如下：
本申请的荧光探针，是一种高选择性、低检测下线、较大的伪斯托克斯位移以及比率远红外ONOO‑荧光探针，能满足医学能对ONOO‑引起的病变及时诊断的需求。
Description
技术领域
本发明属于离子检测技术领域，具体是涉及到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。
背景技术
过氧亚硝酸酰阴离子(ONOO-)是体内最重要的活性氧之一，在生命活动中起着至关重要的作用。正常浓度的过氧亚硝酸酰阴离子在生理和病理过程中起到信号转导和抗菌作用。然而，由于ONOO-较强的氧化氧化和亲核能力，高浓度的ONOO-对许多生物分子造成损伤，如蛋白质、脂类、DNA等，从而可能影响细胞生长过程以及导致细胞坏死和凋亡。因此发展一种高选择、灵敏检测的方法用于ONOO-的检测具有重要意义。近几年来，发展了许多用于ONOO-检测的方法比如电子顺磁共振法、电化学法、色谱法、化学发光和紫外-可见吸收光谱法荧光探针法等，但是上述检测方法需要大型仪器，耗时耗力，不适用于生物体内ONOO-的原位、实时快速定量分析。
荧光探针法由于具有高灵敏性、实时性、较高的时空分辨性以及可以直接用于生物体中活性物质的检测等优势，广泛用于生物体中ONOO-的检测。然而，目前用于ONOO-检测的荧光探针大多以开-关型以及短发射波长为主，其中开-关型荧光探针易环境条件的改变、光漂白以及样品浓度的变化的影响；短发射波长荧光探针不能对生物体中深层组织检测、对光有较大的损伤以及易受激发光的干扰。因此，上述荧光探针不利于ONOO-的精准检测以及对深层组织ONOO-的检测。另外，较小的伪斯托克斯位移的荧光探针易受到生物体荧光的干扰。
用于细胞内H2O2和ONOO-检测的荧光探针的设计合成与生物分析应用，杨秀娥，2016年山东师范大学硕士论文，第一章2.3节公开了过氧亚硝基阴离子荧光探针的研究进展，有荧光光谱法、紫外可见光谱法、化学发光法、电化学方法、酶学法和免疫组织化学法等，包括如下探针化合物，如：
苯基酰肼类
苯并噻唑(BT)类
以花菁为荧光母体，二价硒为响应基团的探针
以香豆素和半花菁为荧光骨架的比率型探针CHCN
以派洛宁为荧光母体
中国专利申请201810684964.0公开了对称的化合物
发明内容
本发明要解决的技术问题是提供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，具有高选择性，低检测下线以及较大的伪斯托克斯位移。
本发明的内容包括一种过氧亚硝酰阴离子比率远红外荧光探针，结构式如下：
一种过氧亚硝酰阴离子比率远红外荧光探针的制备方法，步骤为：
将化合物ANI和碘乙烷溶于有机溶剂中，搅拌回流，待反应完成后，过滤，得到过氧亚硝酰阴离子比率远红外荧光探针，所述化合物ANI的结构为：
所述化合物ANI和碘乙烷的摩尔比为1:2；所述有机溶剂为无水乙腈；所述化合物ANI和无水乙腈的重量比为10-15:1。
具体的反应过程为：
所述化合物ANI的制备方法为，将化合物AN和二苯基次磷酰氯溶于三乙胺和有机溶剂(优选为无水四氢呋喃)中，搅拌，反应得到化合物ANI；化合物AN的结构为：
所述化合物AN、二苯基次磷酰氯和三乙胺的摩尔比为1:1.5:1.5。
本申请还提供一种过氧亚硝酰阴离子比率远红外荧光探针在检测过氧亚硝酰阴离子中的应用。
本发明的有益效果是，本申请的荧光探针，是一种高选择性、低检测下线、较大的伪斯托克斯位移以及比率远红外ONOO-荧光探针，能满足医学能对ONOO-引起的病变及时诊断的需求。
本申请以异佛尔酮吡啶盐为荧光基团，二苯基次磷酰酯为识别基团，探针ANI-DP自身由于羟基被保护，在535nm处有荧光发射。当加入过氧亚硝酰阴离子时，过氧亚硝酰阴离子与二苯基次磷酰酯发生亲核加成反应，二苯基次磷酰酯脱去，生成酚羟基，荧光红移至628nm。
相比于本发明人先前申请的荧光探针，即：
其中，R基团为-H、-OH、-Cl或-OCH3；本申请的反应机理不同，本申请属于分子内质子转移(ICT)机理，属于比率远红外荧光探针，伪斯托克斯位移较大，不易受环境条件的改变、光漂白以及样品浓度变化的影响。
本申请的荧光探针对过氧亚硝酰阴离子检测下线比较低，为13.3nM，说明该探针比较灵敏。
附图说明
图1为探针化合物ANI-DP的1H NMR图谱。
图2为探针化合物ANI-DP的13C NMR图谱。
图3为探针化合物ANI-DP加入(带竖直线的线条)和不加ONOO-的紫外可见光吸收光谱图。
图4为探针化合物ANI-DP与ONOO-作用前后的荧光强度变化图。
图5为本发明荧光探针的荧光强度(I628/I535)比值和ONOO-浓度线性关系图。
图6为探针化合物ANI-DP与ONOO-作用前后荧光强度(I628/I535)比值随pH的变化图。
图7为探针化合物ANI-DP与ONOO-作用前后荧光强度(I628/I535)比值随时间的变化图。
图8为探针化合物ANI-DP与不同的生物活性物种作用后的荧光强度(I628/I535)比值变化图。
(1)ANI-DP，(2)Zn2+，(3)Fe3+，(4)K+，(5)Al3+，(6)Cu2+，(7)NO2 -，(8)NO3 -，(9)F-，(10)Cl-，(11)Br-，(12)CO3 2-，(13)H2PO4 -，(14)AcO-，(15)Cys，(16)Hcy，(17)GSH，(18)H2O2，(19)HClO，(20)HO·-，(21)tBuHO·-，(22)ROO·-，(23)O2·-，(24)1O2 -，(25)ONOO-。
具体实施方式
实施例1
将化合物AN(400mg，1mmol)、二苯基次磷酰氯(0.356mg，1.5mmol)和三乙胺(0.21mL)溶于10mL无水四氢呋喃中，溶液搅拌室温反应12小时，待反应完成后，旋干柱层析的产品ANI(0.288mg，产率48％)。
1H NMR(400MHz,DMSO-d6)δ(ppm):8.60(d,J＝6.4Hz,2H),7.94-7.85(m,5H),7.65-7.62(m,4H),7.58-7.53(m,4H)),7.30(d,J＝8.0Hz,2H),7.12(d,J＝8.0Hz,2H),5.53(s,2H),2.51(s,2H),1.01(s,6H)；13C NMR(100MHz,DMSO-d6)δ(ppm):166.5,162.0,154.7,154.1,151.8,150.3,145.3,135.5,133.3,132.0,131.7,130.6,130.4,129.7,129.5,129.4,125.9,124.0,122.1,121.5,121.4,117.3,116.5,98.2,97.5。
将化合物ANI(600mg，1mmol)和碘乙烷(940mg，2mmol)溶于5mL无水乙腈中，溶液搅拌回流反应8小时，待反应完成后，过滤，收集黄色产品470mg，产率62％)。
1H NMR(400MHz，DMSO-d6)δ(ppm):9.12(d，J＝6.8Hz,2H)，8.13(d,J＝6.8Hz,2H)，7.94-7.85(m,5H)，7.66-7.62(m,4H)，7.57-7.54(m,4H))，7.30(d,J＝8.0Hz,2H)，7.14(d,J＝6.0Hz,2H)，5.59(s,2H)，4.62(q,2H)，2.66(s,2H)，2.49(s,2H)，1.55(t,3H)，1.02(s,6H)；13C NMR(100MHz,DMSO-d6)δ(ppm):167.0,164.0,161.6,155.8,154.6,152.1,151.8,144.9,135.8,133.4,132.0,131.9,131.7,130.3,129.5,127.8,125.5,123.9,121.4,99.7,64.5,56.3,44.8,31.9,28.1,16.7。具体如图1-2所示。
总反应式为：
实施例2
一、探针ANI-DP对ONOO-响应的吸收光谱和荧光光谱
将探针化合物ANI-DP溶于DMSO中，配置成1mmol/L的溶液。向移取的探针ANI-DP溶液中加入DMSO和PBS缓冲液(pH＝7.4)，配置成10μM(DMSO：PBS＝10:90，V/V)的待测溶液。向待测溶液加入ONOO-，用紫外光谱和荧光光谱仪测试其荧光发射光谱的变化情况。由图3可知，探针ANI-DP在388nm处有强的吸收峰，加入ONOO-之后，探针ANI-DP在451nm处有出现了新的吸收峰。由图4可知，探针ANI-DP自身在535nm处有发射峰，加入ONOO-之后，探针在628nm处出现了强荧光发射峰，并且该发射峰强度随着ONOO-浓度的增加而增强，而535nm处发射峰则逐渐减弱。
该探针ANI-DP可以对ONOO-实现比率远红外检测，其伪斯托克斯位移可达到177nm。由图5通过计算可得测下线为13.3nM(计算公式为3σ/k，其中σ为空白样品连续测定多次的标准偏差，k为标准曲线斜率)。
二、pH值变化对探针ANI-DP与ONOO-作用的荧光强度(I554/I473)比值影响
在不同的pH条件下，测试了探针ANI-DP对ONOO-的荧光强度比值(I628/I535)变化。由图6可知，pH为2-4时，探针ANI-DP的荧光强度比值(I628/I535)逐渐增强，pH为4-10时，探针ANI-DP的荧光强度比值(I628/I535)达到最佳，并保持不变。因此，探针ANI-DP可在生理pH条件下实现对ONOO-的检测。
三、探针ANI-DP对ONOO-的时间研究
为了考察探针ANI-DP对ONOO-响应的时间，测试了探针ANI-DP对ONOO-的荧光强度比值(I628/I535)随时间变化情况。由图7可知，在探针ANI-DP溶液加入ONOO-后，荧光强度比值(I628/I535)随时间的变化而逐渐增强，荧光强度比值(I628/I535)在20分钟内达到最大，说明探针ANI-DP可实时检测ONOO-。
四、探针ANI-DP对ONOO-的选择性研究
在探针ANI-DP溶液中分别加入不同的生物活性物种，分别测试了探针ANI-DP的荧光强度比值(I628/I535)变化情况。由图8可知，在探针ANI-DP溶液中加入ONOO-后，荧光强度比值(I628/I535)发生了明显的改变，而加入其它生物活性物种，荧光强度比值(I628/I535)几乎没有发生变化，说明探针ANI-DP能够选择性的实现ONOO-检测。
Claims (6)
1.一种过氧亚硝酰阴离子比率远红外荧光探针，其特征是，结构式如下：
2.一种如权利要求1所述的过氧亚硝酰阴离子比率远红外荧光探针的制备方法，其特征是，步骤为：
将化合物ANI和碘乙烷溶于有机溶剂中，搅拌回流，待反应完成后，过滤，得到过氧亚硝酰阴离子比率远红外荧光探针，所述化合物ANI的结构为：
3.如权利要求2所述的制备方法，其特征是，所述化合物ANI和碘乙烷的摩尔比为1:2。
4.如权利要求2或3所述的制备方法，其特征是，所述有机溶剂为无水乙腈。
5.如权利要求3所述的制备方法，其特征是，所述化合物ANI和无水乙腈的重量比为10-15:1。
6.一种如权利要求1所述的过氧亚硝酰阴离子比率远红外荧光探针在非疾病的诊断中检测过氧亚硝酰阴离子中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010745792.0A CN111848671B (zh) | 2020-07-29 | 2020-07-29 | 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 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010745792.0A CN111848671B (zh) | 2020-07-29 | 2020-07-29 | 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 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111848671A CN111848671A (zh) | 2020-10-30 |
| CN111848671B true CN111848671B (zh) | 2022-06-21 |
Family
ID=72946746
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010745792.0A Active CN111848671B (zh) | 2020-07-29 | 2020-07-29 | 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 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111848671B (zh) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112574252B (zh) * | 2020-11-24 | 2021-08-27 | 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 |
| CN113945552B (zh) * | 2021-10-20 | 2024-08-13 | 延边大学 | 一种活细胞线粒体中还原型谷胱甘肽的浓度测定方法 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107652968A (zh) * | 2017-11-06 | 2018-02-02 | 北京理工大学 | 一种过氧化亚硝酰荧光探针及其制备方法和应用 |
| 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 |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5228190B2 (ja) * | 2005-11-14 | 2013-07-03 | 国立大学法人 東京大学 | パーオキシナイトライト蛍光プローブ |
| GB201403470D0 (en) * | 2014-02-27 | 2014-04-16 | Univ Bath | Method |
-
2020
- 2020-07-29 CN CN202010745792.0A patent/CN111848671B/zh active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107652968A (zh) * | 2017-11-06 | 2018-02-02 | 北京理工大学 | 一种过氧化亚硝酰荧光探针及其制备方法和应用 |
| 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 |
Non-Patent Citations (3)
| Title |
|---|
| A fluorogenic and red-shifted diphenyl phosphinate-based probe for selective peroxynitrite detection as demonstrated in fixed cells;Sandip V. M.et al.;《New J.Chem.》;20170904;第41卷;第11934-11940页 * |
| Rational construction of a novel ratiometric far-red fluorescent probe with excellent water solubility for sensing mitochondrial peroxynitrite;Gu Biao et al.;《Sensors and Actuators: B. Chemical》;20210606;第344卷;第130246-130254 页 * |
| Synthesis and biological activity of substituted benzo[b]thiophenes, benzothieno[2,3-d]pyridines, benzothieno[2,3-d]pyrimidines and benzothieno[2,3-d]pyrazoles;Desai J.M. et al.;《Indian Journal of Chemistry, Section B: Organic Chemistry Including Medicinal Chemistry》;19971231;第36B卷(第8期);第668-674页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111848671A (zh) | 2020-10-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wei et al. | NBD-based colorimetric and fluorescent turn-on probes for hydrogen sulfide | |
| Gunnlaugsson et al. | Novel sodium-selective fluorescent PET and optically based chemosensors: towards Na+ determination in serum | |
| Tan et al. | Functionalized lanthanide coordination polymer nanoparticles for selective sensing of hydrogen peroxide in biological fluids | |
| Huo et al. | The synthesis, characterization of three isomers of rhodamine derivative and their application in copper (II) ion recognition | |
| CN108752377B (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 | |
| 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 | |
| CN109971464B (zh) | 一种基于氧杂蒽和香豆素的区分过氧化亚硝基和次氯酸根荧光探针的制备方法 | |
| CN110330965B (zh) | 一种含脲结构的六元螺环罗丹明pH荧光指示剂及其应用 | |
| CN111116696A (zh) | 一种三磷酸腺苷近红外纳米荧光探针的制备和应用 | |
| Zhu et al. | Near-infrared cyanine-based sensor for Fe 3+ with high sensitivity: its intracellular imaging application in colorectal cancer cells | |
| Ding et al. | The design and synthesis of two imidazole fluorescent probes for the special recognition of HClO/NaHSO 3 and their applications | |
| CN110229165A (zh) | 上转换荧光探针罗丹明衍生物及其应用 | |
| Yoon et al. | Synthesis and evaluation of self-calibrating ratiometric viscosity sensors | |
| CN109021000B (zh) | 一种检测过氧化氢的荧光探针、合成方法和应用 | |
| CN110357896B (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 | |
| CN115651006B (zh) | 一种具有大斯托克斯位移的过氧化氢比率型近红外荧光探针及其制备方法与应用 | |
| KR101079315B1 (ko) | 신규 로다민 유도체 및 이를 포함하는 수은 이온 검출용 센서 | |
| CN111808607B (zh) | 一种检测过氧亚硝酰阴离子的荧光探针、制备方法和应用 | |
| CN114014891B (zh) | 一种嘌呤环修饰的荧光探针及其在检测钴离子上的应用 | |
| Liang et al. | A highly selective and sensitive ratiometric fluorescent probe for pH measurement based on fluorescence resonance energy transfer | |
| CN114773305A (zh) | 一种2-2环芳吡喃酮pH荧光比率探针的制备方法及应用 | |
| CN113061139A (zh) | 一种含氨基脲结构的六元螺环罗丹明荧光探针及其制备方法和应用 | |
| Şenkuytu et al. | Novel probes for selective fluorometric sensing of Fe (II) and Fe (III) based on BODIPY dyes | |
| Zhao et al. | A novel isophorone-based fluorescent probe for recognizing Al3+ and its bioimaging in plants | |
| Yao et al. | A novel “on–off–on” near-infrared fluorescent probe for Cu 2+ and S 2− continuous detection based on dicyanoisoflurone derivatives, and its application in bacterial imaging |
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 |