CN113603722B - 一种极性荧光探针及其制备方法和应用 - Google Patents
一种极性荧光探针及其制备方法和应用 Download PDFInfo
- Publication number
- CN113603722B CN113603722B CN202111027991.9A CN202111027991A CN113603722B CN 113603722 B CN113603722 B CN 113603722B CN 202111027991 A CN202111027991 A CN 202111027991A CN 113603722 B CN113603722 B CN 113603722B
- Authority
- CN
- China
- Prior art keywords
- probe
- polarity
- fluorescent probe
- change
- preparation
- 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 20
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 239000000523 sample Substances 0.000 claims abstract description 47
- 239000002904 solvent Substances 0.000 claims abstract description 18
- 239000000126 substance Substances 0.000 claims abstract description 8
- 239000003153 chemical reaction reagent Substances 0.000 claims description 2
- 230000008859 change Effects 0.000 abstract description 13
- 230000007613 environmental effect Effects 0.000 abstract description 8
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 24
- 239000000243 solution Substances 0.000 description 17
- 238000006243 chemical reaction Methods 0.000 description 15
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 12
- 238000004440 column chromatography Methods 0.000 description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 8
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 8
- 238000001035 drying Methods 0.000 description 8
- 238000003756 stirring Methods 0.000 description 8
- 229940125782 compound 2 Drugs 0.000 description 7
- 229940126214 compound 3 Drugs 0.000 description 7
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 6
- 238000005160 1H NMR spectroscopy Methods 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- MZRVEZGGRBJDDB-UHFFFAOYSA-N N-Butyllithium Chemical compound [Li]CCCC MZRVEZGGRBJDDB-UHFFFAOYSA-N 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 4
- RNHDAKUGFHSZEV-UHFFFAOYSA-N 1,4-dioxane;hydrate Chemical compound O.C1COCCO1 RNHDAKUGFHSZEV-UHFFFAOYSA-N 0.000 description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 229960000583 acetic acid Drugs 0.000 description 4
- 239000012267 brine Substances 0.000 description 4
- 230000001413 cellular effect Effects 0.000 description 4
- 239000012362 glacial acetic acid Substances 0.000 description 4
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 4
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 4
- JNRLEMMIVRBKJE-UHFFFAOYSA-N 4,4'-Methylenebis(N,N-dimethylaniline) Chemical compound C1=CC(N(C)C)=CC=C1CC1=CC=C(N(C)C)C=C1 JNRLEMMIVRBKJE-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- IMDXZWRLUZPMDH-UHFFFAOYSA-N dichlorophenylphosphine Chemical compound ClP(Cl)C1=CC=CC=C1 IMDXZWRLUZPMDH-UHFFFAOYSA-N 0.000 description 3
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 230000003834 intracellular effect Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- UGNWTBMOAKPKBL-UHFFFAOYSA-N tetrachloro-1,4-benzoquinone Chemical compound ClC1=C(Cl)C(=O)C(Cl)=C(Cl)C1=O UGNWTBMOAKPKBL-UHFFFAOYSA-N 0.000 description 3
- USEXQPWLCGBYNT-UHFFFAOYSA-N 3-bromo-n,n-dimethylaniline Chemical compound CN(C)C1=CC=CC(Br)=C1 USEXQPWLCGBYNT-UHFFFAOYSA-N 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 229940125904 compound 1 Drugs 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000000295 emission spectrum Methods 0.000 description 2
- 238000002189 fluorescence spectrum Methods 0.000 description 2
- 239000003208 petroleum Substances 0.000 description 2
- 230000035790 physiological processes and functions Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000010791 quenching Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 229920006395 saturated elastomer Polymers 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 206010019695 Hepatic neoplasm Diseases 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000012292 cell migration Effects 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- FRIPRWYKBIOZJU-UHFFFAOYSA-N fluorone Chemical compound C1=CC=C2OC3=CC(=O)C=CC3=CC2=C1 FRIPRWYKBIOZJU-UHFFFAOYSA-N 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- -1 phenylphosphoryloxypyridine Chemical class 0.000 description 1
- 208000030761 polycystic kidney disease Diseases 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011897 real-time detection Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 238000009987 spinning Methods 0.000 description 1
- 239000011550 stock solution Substances 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
- 238000012546 transfer Methods 0.000 description 1
- 210000004926 tubular epithelial cell Anatomy 0.000 description 1
- 238000002371 ultraviolet--visible spectrum Methods 0.000 description 1
- 230000028973 vesicle-mediated transport Effects 0.000 description 1
- 238000012800 visualization 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
- C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
- C07F9/6564—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms
- C07F9/6568—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms having phosphorus atoms as the only ring hetero atoms
- C07F9/65685—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having phosphorus atoms, with or without nitrogen, oxygen, sulfur, selenium or tellurium atoms, as ring hetero atoms having phosphorus atoms as the only ring hetero atoms the ring phosphorus atom being part of a phosphine oxide or thioxide
-
- 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
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1096—Heterocyclic compounds characterised by ligands containing other heteroatoms
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Optics & Photonics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
本发明属于荧光探针领域，具体涉及一种极性荧光探针及其制备方法和应用。为解决通过荧光强度变化来检测极性的荧光探针在实际应用时易受探针浓度及环境因素干扰等问题，本发明提供了一种通过波长变化来检测极性的荧光探针。该探针结构中包含着D‑A‑D结构，其发射波长随着体系极性的增大而逐渐红移，发射波长与0.020‑0.287的极性范围呈良好的线性关系。因此，该探针为准确检测化学溶剂或者细胞内的极性变化提供了一个潜在的工具。
Description
技术领域
本发明属于荧光探针领域，具体涉及一种极性荧光探针及其制备方法和应用。
背景技术
极性不仅是化学和化工领域中一个非常重要的化学参数，也是细胞微环境的重要参数之一。细胞极性是细胞中某些胞质成分按照一定空间顺序的不均等分布，从而形成各种细胞内容物的浓度梯度，正是由于细胞极性的存在导致了细胞的不对称分裂。在生物体系中，极性在调节细胞迁移、增殖、分化、囊泡转运、激素分布和免疫反应等关键生理过程中发挥着关键作用。例如，蛋白质可以通过改变构象调节其内部极性，来控制酶促反应速率；小管上皮细胞的极性是小管吸收、分泌排泄和交换等功能的基础等。细胞内极性微环境的异常还与某些代谢性疾病的发生和发展密切相关，如常染色体显性遗传性多囊肾、肝硬化及肿瘤等。因此，开发能检测细胞极性的荧光探针对于研究蛋白质的生理功能或防治疾病等均具有重要意义。
鉴于高敏感性、可视化、非侵袭性、无辐射、实时检测、低成本等特点，荧光技术已经是生物学、医学、药学等领域不可或缺的研究手段，在食品与药品安全检测、环境监测、医学诊断等方面发挥了重要作用。近年来，极性荧光探针因具有检测灵敏性高，生物兼容性好等优点，被认为是检测细胞内极性变化的理想工具。目前报道的极性荧光探针的常用设计策略是基于分子内电荷转移(ICT)机理，即探针结构中包含着推/拉电子体系，当体系的极性发生改变时会引起探针荧光强度或者波长的改变。基于荧光强度改变的极性探针在实际应用时可能会受到探针浓度、生物自发荧光或者环境因素的影响，而基于波长改变的极性探针则可以有效避免上述问题。针对这一问题，本发明开发了一种基于苯基磷氧吡啰红酮荧光团的新型极性荧光探针，该探针可以通过波长变化来检测极性的改变，不受探针浓度、生物自发荧光或者环境因素的、干扰，因此，该探针为准确检测化学溶剂或者细胞内的极性变化提供了一个潜在的工具。
发明内容
针对上述问题本发明提供了一种极性荧光探针及其制备方法和应用，该探针结构中包含着D-A-D结构，其发射波长随着体系极性的增大而逐渐红移，探针的发射波长与0.020-0.287的极性范围呈良好的线性关系。因此，该探针有望成为准确检测化学溶剂或者细胞内极性的重要工具。
为了达到上述目的，本发明采用了下列技术方案：
一种极性荧光探针，所述探针的结构式为：
一种极性荧光探针的制备方法，其特征在于，包括以下步骤：
(1)将3-溴-N,N-二甲基苯胺(化合物1)溶解在冰醋酸中，向上述溶液中逐滴滴入福尔马林溶液，进行搅拌反应，反应结束后，将冰醋酸旋干，得到的固体经饱和NaHCO3溶液中和、萃取、洗涤、干燥和柱色谱分离后得到4，4’-亚甲基双(N，N-二甲基苯胺)(化合物2)。
(2)将4，4’-亚甲基双(N，N-二甲基苯胺)(化合物2)溶解在无水THF中，反应体系在无水无氧条件下冷却至-78℃，向上述溶液中逐滴加入正丁基锂，维持此温度搅拌；将苯基二氯膦溶解在无水THF中，并将此溶液逐滴滴入上述反应液中，反应液升至室温并反应过夜；反应结束后加盐酸溶液猝灭反应，反应液经萃取、洗涤、干燥和柱色谱分离后得到二氢苯基磷吡啰红(化合物3)。
(3)将二氢苯基磷吡啰红(化合物3)溶解在THF中，逐渐加入四氯苯醌，室温搅拌后旋干溶剂，得到的固体经柱色谱分离后得到所述极性荧光探针PPyK。
进一步，所述步骤(1)中福尔马林溶液的质量分数为37％，所述3-溴-N,N-二甲基苯胺与福尔马林溶液的摩尔体积比为5mol：1mL。
进一步，所述步骤(1)中搅拌反应的温度为90℃，时间为1h，所述萃取用二氯甲烷，所述洗涤用盐水，所述干燥用无水硫酸镁，所述柱色谱分离展开剂乙酸乙酯：正己烷的体积比为1：30。
进一步，所述步骤(2)中4，4’-亚甲基双(N，N-二甲基苯胺)、正丁基锂与苯基二氯膦的摩尔比为1:2:1。
进一步，所述步骤(2)中搅拌的时间为1.5h，所述盐酸溶液的浓度为2mol/L，所述萃取用二氯甲烷，所述洗涤用盐水，所述干燥用无水硫酸镁，所述柱色谱分离展开剂二氯甲烷：石油醚的体积比为2：5。
进一步，所述步骤(3)中二氢苯基磷吡啰红与四氯苯醌的摩尔比为1:3。
进一步，所述步骤(3)中室温搅拌的时间为15分钟，所述柱色谱分离展开剂二氯甲烷：甲醇体积比为20：1。
一种极性荧光探针的应用，在制备检测化学溶剂或者细胞内极性试剂的应用。
与现有技术相比本发明具有以下优点：
(1)探针合成步骤简单，原料便宜；
(2)探针通过波长变化来检测极性的改变，不受探针浓度及环境因素干扰；
(3)该探针生物兼容性好，有望应用于活体中。
附图说明
图1为化合物2的1H NMR图(CDCl3,600MHz)；
图2为化合物2的13C NMR图(CDCl3,150MHz)；
图3为化合物2的HRMS图；
图4为化合物3的1H NMR图(CDCl3,600MHz)；
图5为化合物3的13C NMR图(CDCl3,150MHz)；
图6为化合物3的HRMS图；
图7为探针PPyK的1H NMR图(CDCl3,600MHz)；
图8为探针PPyK的13C NMR图(CDCl3,150MHz)；
图9为探针PPyK的HRMS图；
图10为探针PPyK在不同溶剂中的紫外可见吸收光谱图。
图11为探针PPyK在不同溶剂中归一化的荧光光谱图；
图12(A)为探针PPyK在不同比例的水和1，4-二氧六环中归一化的荧光光谱图；图12(B)为探针的最大发射波长与溶剂极性的线性关系图。
具体实施方式
实施例1
一种极性荧光探针，其特征在于，所述探针的结构式为：
一种所述极性荧光探针的制备方法，其特征在于，包括以下步骤：
(1)将化合物1(5g，25mmol)溶解在冰醋酸(40mL)中，向上述溶液中逐滴滴入37％的福尔马林溶液(5mL)，混合物在90℃下搅拌反应1h；反应结束后，将冰醋酸旋干，得到的固体经饱和NaHCO3中和、二氯甲烷(3×100mL)萃取、盐水洗涤、无水硫酸镁干燥和柱色谱分离(乙酸乙酯：正己烷＝1：30)后得到化合物2(6.18g，60.0％)。
1H NMR(600Hz,CDCl3)δ7.20(m,7H),7.06(d,J＝12.0Hz,2H),6.70(d,J＝8.4Hz,2H),3.63(s,2H),2.90(s,12H)；13C NMR(150MHz,CDCl3)δ152.0,151.9,141.4,141.3,138.2,135.1,135.0,134.2,131.4,131.1,131.0,130.6,121.8,121.5,116.3；ESI-MS:[M+H]+calcd for413.01,Found 413.00.
(2)将化合物2(1.23g，3mmol)溶解在无水THF(10mL)中，反应体系在无水无氧条件下冷却至-78℃，向上述溶液中逐滴加入正丁基锂(6mmol)，维持此温度搅拌1.5h；将苯基二氯膦(0.54g，3mmol)溶解在无水THF中(5mL)，并将此溶液逐滴滴入上述反应液中，反应液缓慢升至室温并反应过夜；反应结束后加盐酸(2mol/L，10mL)猝灭反应，反应液经二氯甲烷(3×100mL)萃取、盐水洗涤、无水硫酸镁干燥和柱色谱分离(二氯甲烷：石油醚＝2：5)后得到化合物3(0.571g，52.9％)。
1H NMR(600Hz,CDCl3)δ7.20(m,7H),7.06(d,J＝12.0Hz,2H),6.70(d,J＝8.4Hz,2H),3.63(s,2H),2.90(s,12H)；13C NMR(150MHz,CDCl3)δ152.0,151.9,141.4,141.3,138.2,135.1,135.0,134.2,131.4,131.1,131.0,130.6,121.8,121.5,116.3；ESI-MS:[M+H]+calcd for 361.18,Found 361.18.
(3)将化合物3(0.36g，1mmol)溶解在THF(20mL)中，敞口逐批加入四氯苯醌(0.738g，3mmol)，室温搅拌15分钟后旋干溶剂，得到的固体经柱色谱分离(二氯甲烷：甲醇＝20：1)后得到探针PPyK(0.334g，85.6％)。
1H NMR(600Hz,CDCl3)δ8.33(d,J＝9.0Hz,1H),8.31(d,J＝9.0Hz,1H),7.60(m,2H),7.38(m,1H),7.32(m,2H),7.15(dd,J1＝2.4Hz,J2＝9.0Hz,2H),6.87(dd,J1＝2.4Hz,J2＝9.6Hz,2H),3.07(s,12H)；13C NMR(150MHz,CDCl3)δ183.3,183.2,155.5,155.4,138.4,137.9,137.7,137.3,134.2,134.1,133.3,133.2,131.5,131.4,127.4,127.3,117.5,114.8,114.7,42.9；ESI-MS:calcd for 391.16,Found391.16.
实施例2
1.测试溶液配制
将探针用乙腈配成2mM的储存液，随后用不同溶剂或者不同比例的水和1，4-二氧六环溶液稀释至测试浓度。
2.光物理性质研究
将10μL的探针分别加到2mL不同极性的溶剂中，从紫外-可见吸收光谱可以看出，探针有两个吸收峰，分别在350nm处和430nm左右，且短波的吸收峰几乎不随溶剂极性的改变而发生位移，只有长波处的吸收峰会随着溶剂极性的增大发生红移(图10)，因此，我们仅检测探针的长波长随溶剂极性的变化。随后，选取400nm为激发波长，测试探针在不同极性的溶剂的发射光谱，如图11所示，探针的发射波长随溶剂极性的增大而逐渐红移。上述结果说明，本发明提供的探针是一个对极性敏感的探针，即探针的发射波长随着环境极性的增大而红移，与传统荧光增强型极性探针相比，该探针不受探针浓度以及环境因素干扰。
3.探针波长与极性的关系研究
为模拟不同的极性，测试了探针在不同比例的水和1，4-二氧六环中的发射光谱，如图12(A)所示，探针的发射波长随水比例的增大而逐渐红移。为研究探针的发射波长与环境极性的定量关系，绘制了探针波长与不同比例的水和1，4-二氧六环对应的极性的线性关系图，如图12(B)所示，探针的波长与0.020-0.287(0-40％含水量)的极性范围呈良好的线性关系，线性相关系数R＝0.999。因此，可以根据本探针在待检测溶剂中的波长，来精确检测0.020-0.287的极性范围。
以上所述仅为本发明的较佳实施例而已，并不用以限制本发明，凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
Claims (1)
1.一种极性荧光探针的应用，其特征在于，在制备检测化学溶剂极性的试剂中的应用，所述化学溶剂极性的范围为：0.020-0.287；
所述探针的结构式为：
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111027991.9A CN113603722B (zh) | 2021-09-02 | 2021-09-02 | 一种极性荧光探针及其制备方法和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111027991.9A CN113603722B (zh) | 2021-09-02 | 2021-09-02 | 一种极性荧光探针及其制备方法和应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113603722A CN113603722A (zh) | 2021-11-05 |
| CN113603722B true CN113603722B (zh) | 2022-05-31 |
Family
ID=78309999
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111027991.9A Active CN113603722B (zh) | 2021-09-02 | 2021-09-02 | 一种极性荧光探针及其制备方法和应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113603722B (zh) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114716479B (zh) * | 2022-05-05 | 2024-01-05 | 广州青苗新材料科技有限公司 | 一种具有热活化延迟荧光性质的膦氧类化合物及其制备与应用 |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110003173A (zh) * | 2019-04-26 | 2019-07-12 | 安徽大学 | 一种基于咔唑的双光子极性荧光探针及其制备方法和用途 |
| CN110028956A (zh) * | 2019-05-24 | 2019-07-19 | 济南大学 | 一种检测脂滴极性的荧光探针及其应用 |
| CN110156688A (zh) * | 2019-05-22 | 2019-08-23 | 济南大学 | 一种靶向内质网检测极性的荧光探针及其应用 |
| CN112341453A (zh) * | 2020-11-05 | 2021-02-09 | 山西大学 | 一种基于香豆素的荧光探针及其制备方法和应用 |
-
2021
- 2021-09-02 CN CN202111027991.9A patent/CN113603722B/zh active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110003173A (zh) * | 2019-04-26 | 2019-07-12 | 安徽大学 | 一种基于咔唑的双光子极性荧光探针及其制备方法和用途 |
| CN110156688A (zh) * | 2019-05-22 | 2019-08-23 | 济南大学 | 一种靶向内质网检测极性的荧光探针及其应用 |
| CN110028956A (zh) * | 2019-05-24 | 2019-07-19 | 济南大学 | 一种检测脂滴极性的荧光探针及其应用 |
| CN112341453A (zh) * | 2020-11-05 | 2021-02-09 | 山西大学 | 一种基于香豆素的荧光探针及其制备方法和应用 |
Non-Patent Citations (3)
| Title |
|---|
| A near-infrared fluorophore for live-cell super-resolution microscopy of cellular protein;Lukinavičius, G.等;《Nat. Chem.》;20131231;132-139 * |
| Effects of Amino Group Substitution on the Photophysical Properties and Stability of Near-Infrared Fluorescent P-Rhodamines;Grzybowski, Marek等;《Chemistry - A European Journal》;20200603;7912-7917 * |
| Rational Design of a Near-infrared Fluorescence Probe for Ca2+ Based on Phosphorus-substituted Rhodamines Utilizing Photoinduced Electron Transfer;Takahashi, Shodai等;《Chemistry - An Asian Journal》;20200107;524-530 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113603722A (zh) | 2021-11-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110172337B (zh) | 一种苯并噻唑衍生物荧光探针及其制备方法和应用 | |
| US11320428B2 (en) | Probe for dual-mode bio-imaging | |
| CN108440475A (zh) | 一种区分不同极性脂滴的比率型荧光探针及其制备方法和应用 | |
| CN107089937A (zh) | 线粒体靶向测定粘度的荧光探针及其制备方法和应用 | |
| CN103214875B (zh) | 一类以荧光素类似物为母体的荧光染料的合成和应用 | |
| CN108219780B (zh) | 一种近红外荧光探针及其制备方法和应用 | |
| CN113603722B (zh) | 一种极性荧光探针及其制备方法和应用 | |
| CN105154065B (zh) | 一种快速专一性识别羟基自由基的荧光探针及其制备方法和应用 | |
| CN108864058A (zh) | 一类氧杂蒽酮类荧光染料及应用 | |
| CN108774226B (zh) | 一种用于检测银离子的荧光探针及其制备方法与应用 | |
| CN105111415A (zh) | 一种侧链含萘基团的聚噻吩衍生物及其制备方法和应用 | |
| CN104650610A (zh) | 一种不对称近红外bodipy荧光染料及其制备和应用 | |
| CN103937487A (zh) | 一种磷光氟离子探针及其制备和应用 | |
| CN109456250A (zh) | 热激活延迟荧光(tadf)纳米探针及其制备方法和在生物成像中的应用 | |
| CN109369684A (zh) | 一类电子供体-受体-供体荧光分子及其制备方法和应用 | |
| CN108191880B (zh) | 一种具有咔唑-罗丹明杂化结构的荧光染料制备方法和应用 | |
| CN111303111A (zh) | 一类新型黄菁近红外二区染料、制备方法及荧光成像应用 | |
| CN108727277B (zh) | 一种单线态氧高度选择性聚集诱导化学发光探针及其制备方法和应用 | |
| Renno et al. | Near‐Infrared Squaraine Dyes as Bright Fluorescent Probes: A Structure–Activity Photophysical Investigation in Liposomes | |
| CN106366041B (zh) | 一种连续识别钯离子、co的荧光探针及应用 | |
| CN106699778B (zh) | 一种近红外荧光染料的制备方法及其应用 | |
| CN112480025B (zh) | 具有聚集诱导发光功能的化合物及其制备方法和应用 | |
| US10029996B2 (en) | Class of cyano-substituted asymmetric cyanine dyes, synthesizing method and application thereof | |
| CN112341453A (zh) | 一种基于香豆素的荧光探针及其制备方法和应用 | |
| Guo et al. | One-pot synthesis and applications of two asymmetrical benzoxanthene dyes |
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 |